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All hotels in Galveston, Texas, United States of America
1 room, 2 adults
8216 Harborside Dr, Galveston, TX, 77554, United States of America, 855-239-9481
Howard Johnson Express Inn - Galveston Texas, Galveston
Lowest price$68
available on 06/30/16
Best Price Guarantee
Convenient to Moody Gardens
- Free continental breakfast, free WiFi, and free parking
Main amenities
- 46 guestrooms
- Air conditioning
- Daily housekeeping
For families
- Rollaway/extra beds available
- Television
- Daily housekeeping
- Iron/ironing board
- Hair dryer
- Coffee/tea maker
What's around
- Galveston Schlitterbahn Waterpark (4.2 mi / 6.8 km)
- Moody Gardens (4.2 mi / 6.8 km)
- Lone Star Flight Museum (4.4 mi / 7.2 km)
- Galveston Island Railroad Museum (4.5 mi / 7.2 km)
- Texas Seaport Museum (4.8 mi / 7.8 km)
See all 261 Hotels.com reviews
Fantastic service
Great location
Beach is poor
TripAdvisor Traveller Rating: 3
From 211 reviews
Howard Johnson Express Inn - Galveston Texas, Galveston
Hotel Info: 866-539-8117from $68
- Standard Room, 2 Queen Beds, Smoking
- Standard Room, 1 King Bed, Accessible
- Standard Room, 2 Queen Beds
- Standard Room, 1 King Bed
- Standard Room, 1 King Bed, Smoking
Also popular with travelers from the USA
At a glance
Key facts
Hotel size
- This hotel has 46 rooms
- This hotel is arranged over 2 floors
- Check-in time 3 PM-Midnight
- Check-out time is Noon
Required at check in
- Credit card or cash deposit required
- Government-issued photo ID required
- Minimum check-in age is 21
Travelling with others
- No cribs (infant beds)
- Pets not allowed
Extra people
- Roll-away beds available, charges may apply*
- Free WiFi in public areas
- Free WiFi in rooms
- Free self parking
- RV/bus/truck parking
Nearby Two Star Galveston hotels
In the hotel
Food and drink
- Free continental breakfast daily
- Accessible bathroom
In the room
Home comforts
- Air conditioning
- Coffee/tea maker
- Iron/ironing board
Freshen up
- Hair dryer
Be entertained
- Cable TV channels
Stay connected
- Desk
- Free WiFi
- Free local calls
- Daily housekeeping
- Access via exterior corridors
Top Galveston hotels
Recently reviewed hotels in Galveston
- Scottish Inns GalvestonFrom
"The only downside to my stay where the shower water pressure or lack thereof. Otherwise I would stay there again I hope they fix the water pressure because I do enjoy bathing."Jun 14, 2016
- Hampton Inn and Suites Houston/League CityFrom
"Check in was very easy. Front desk people were very friendly and great service. Our family with 2 kids had a pleasant stay. Very good breakfast in the morning. We will come back…"Jun 14, 2016
- SpringHill Suites by Marriott Houston NASA/WebsterFrom
"The hotel is located near elevated freeways on two sides with major power lines on a third. Highway dependent fast food restaurants and gas stations abound in the area."Jun 13, 2016
- Microtel Inn & Suites by Wyndham HoustonFrom
"Was comfortable and clesn. I loved it,I will go back."Jun 13, 2016
Nearby hotels
Recent reviews
Good 3.0 /5 from 261 reviews
Howard Johnson Express Inn - Galveston Texas, Galveston
There are definitely better hotels around the same price in the area to choose from. We knew nothing about the area when we booked. Not a great part of town. My husband and I both had a sore back after sleeping on the awful mattress.
Howard Johnson Express Inn - Galveston Texas, Galveston
If you are looking for a cheap and decent place to stay in Galveston, this is definitely an option to consider.
Howard Johnson Express Inn - Galveston Texas, Galveston
Wouldn't stay again.
Hotel needs some work and care.
Howard Johnson Express Inn - Galveston Texas, Galveston
Ok for a night
We wanted a budget hotel to stay in the night before our cruise. While it was not the most comfortable hotel, it was easy to get to the port the following day. The service was excellent and the person working at check in and check out was extremely friendly and helpful. The breakfast area has maybe 4 tables but it worked for us. We didn't sleep well as there are train tracks right behind the hotel and a truck stop gas station on the other side. It was rather alarming waking up to the train horn throughout the night.
Howard Johnson Express Inn - Galveston Texas, Galveston
It was awful, loud noise, trains trucks etc. I was seriously afraid, it was dirty the secondary latch on door was gone, the bedding was dingy and one even had stains! I considering leaving and sleeping in my car!
Is the description of this hotel not correct? Tell us


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Tip for Living: 'Bringing Up Our Children in Light and Truth' offers model for Godly parenting
Most people want to be good parents. They want to raise happy, healthy children and maintain close, loving relationships with them. They want their homes to be filled with peace and love. They begin their journey through parenthood with the highest ideals and the best of intentions.
These "noble aspirations" are often thwarted when parents are confronted with the trials and struggles of daily life, H. Wallace Goddard says, adding that "parents get tired, distracted, frustrated, and overwhelmed." In his recent book "Bringing Up Our Children in Light and Truth" (Walnut Springs Press, $14.99), he suggests the use of basic gospel principles to make the goal of being a good parent a little easier to accomplish.
Goddard believes that parenting "is, above all else, an apprenticeship for godliness" and we cannot be successful without divine help. He likens the process to building a house and offers the following as a "Model for Godly Parenting."
• Footing — Be a flourishing person
Parents must be healthy, balanced individuals. This includes not only mental, emotional and physical health, but spiritual health as well. To become a flourishing person, parents need to show gratitude, share their talents and serve others.
• Foundation — Have compassion
Godly parenting cannot exist without compassion. Goddard defines compassion as the "divine gift of feeling what another person feels even though we do not have his or her life experience." He says that we "simply are not capable of true compassion without heavenly help."
• Body — Nurture and guide
Parents must love, support and encourage their children. They must also teach them about eternal principles and the consequences of their actions. According to Goddard, nurture and guidance are equal partners in parenting. He writes that "guidance of children is not effective in the absence of nurture. And nurture without guidance is indulgence."
• Roof — Have an eternal purpose
Parents must remember that they are not merely trying to get their children through this life. They are also trying to prepare them to serve their Heavenly Father. They have an eternal purpose and should continually seek heavenly guidance to raise their children in "light and truth."
Goddard is a professor of family life for the University of Arkansas Cooperative Extension Service. He and his wife Nancy are members of The Church of Jesus Christ of Latter-day Saints. They have three adult children, 10 grandchildren and have cared for 20 foster children during their 37 years of marriage.
Sandra Nazar lives, writes and blogs in Oklahoma with her husband and five children. She blogs at www.sincerelysandra.net.
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subset: commoncrawl


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Tom Cruise and Katie Holmes Officially Divorced
Tom Cruise and Katie Holmes are officially back on the market again. The parents of 6-year-old Suri get their divorce papers signed off by judge just a month and a half after the actress submitted her filing for legal separation from the actor.
A source tells E! News that the "Top Gun" actor is relieved that the proceedings are finally over. "Tom is very happy and relieved that he is moving forward," the source testifies. "His priority is being a good father and this settlement will make sure that continues."
Tom and Katie, who have a prenup agreement, settle their divorce outside the court. Suri will live primarily with Katie in New York with generous visitation by Tom. The mother-daughter duo currently lives in an apartment located in Manhattan's artsy Chelsea neighborhood.
Other couples splitting on amicable terms are Katy Perry & Russell Brand and Courteney Cox & David Arquette. The former two have already had the union officially dissolved, while the latters haven't but they continue to keep it civil.
More Tom Cruise News
- Video: Lady GaGa and Caitlyn Jenner Among 49 Stars Honoring Orlando Shooting Victims
- Justin Timberlake Is One Grumpy Doll in First 'Trolls' Trailer
- Stephanie March Reveals Botched Boob Job Amid 2014 Bobby Flay Marriage Trouble
- Peta Murgatroyd Confirms She's Pregnant With Maksim Chmerkovskiy's Baby
- Drew Barrymore in Talks to Host New Talk Show
- Kanye West Signs New Partnership With Adidas to Expand Yeezy Brand
- Dennis Quaid's Wife Kimberly Files for Divorce for Second Time
- Chris Brown Rants on Instagram After Falling Out with Manager and Publicist


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Iration Tickets for Sale
Iration is always a great hit with every crowd. We stock all tickets to every Iration show!
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|Jul 7 Thu 8:00 PM
|House Of Blues - San Diego
San Diego, CA
|View Tickets
|Jul 9 Sat 5:00 PM
|Iration, Rebel Souljahz, Katastro & The Steppas
|Pacific Amphitheatre
Costa Mesa, CA
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|Jul 10 Sun 5:00 PM
|Iration, Collie Buddz, Seedless & The Expanders
|Pacific Amphitheatre
Costa Mesa, CA
|View Tickets
|Aug 20 Sat 1:00 PM
|Michael Franti And Spearhead, Iration, Common Kings, Fortunate Youth & Hirie
|Thunder Valley Casino - Amphitheatre
Lincoln, CA
|View Tickets
|Aug 24 Wed 7:30 PM
|Michael Franti and Spearhead & Iration
|Antelope Valley Fair
Lancaster, CA
|View Tickets
|Aug 27 Sat 2:00 PM
|Reggae On The Rocks: 311, Iration, Steel Pulse, Morgan Heritage, The Werks & Judge Roughneck
|Red Rocks Amphitheatre
Morrison, CO
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Who the bloody hell knows what the founded fathers were envisioning or intending?
Fun fact: You weren't alive in the 1700's. None of us here were. What the heck do you know about what they were thinking or what they would be thinking?
It matters because the Amendment was written during a time when the current technology is lightyears beyond anything in their wildest dreams. So a document for 1789 could not have been written that accounted for that kind of wholesale destruction. Applying modern interpretations to a 230ish year old piece of paper doesn't transfer & jive well. Hence the need for loose constructionalism.


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"Justice League": Exploring How Superman Returns (Again)
Film, Comic Books
Conventions | Cox Communications and Comic-Con International will provide free WiFi to the entire downtown area of San Diego from July 8 to July 24, a period that encompasses the 2016 MLB All-Star Game as well as Comic-Con International. However, the WiFi will only be available outside the convention center during Comic-Con. Cox will install 100 hotspots around town, and for the period of Comic-Con will make them available for free to all users. After July 24, the hotspots will be available to Cox customers, and non-subscribers will be allowed one free hour per month. [San Diego Union-Tribune]
Manga | As part of its Cool Japan initiative, the Japanese government plans to build the "Manga National Center" — a museum dedicated to manga, anime and video games — in time for the 2020 Tokyo Olympics and Paralympics. Expected to cost about $90 million, the project will be funded through a mixture of public and private-sector money. [Chicago Tribune]
Passings | Augie Scotto, an artist whose work appeared in Will Eisner's PS magazine, passed away March 15 at age 88. He began his career in 1949, drawing largely crime and Western stories for such early publishers as Eastern Color, Atlas and Charlton. Scotto seems to have left comics for a while around 1953, but returned in 1968 as the penciler for Tower Comics' Dynamo and as an inker for DC Comics until around 1978. [Timely-Atlas-Comics]
Collecting | For the better part of three decades, 78-year-old Ray Brown has been "rescuing" comic books and giving them a good home — namely, his. The South Dakota man estimates his collection includes some 75,000 comics, the bulk of which he purchased from five Rapid City-area stores that went out of business. "They take up a lot of room," he says. "They don't eat anything, though." Brown doesn't read them, however; instead he simply takes pleasure in saving them from the trash bin. He does sell a few on the Internet from time to time, but he's in no hurry to get rid them. [Black Hills Fox]
Legal | The trial began Monday in San Diego for Matthew Pocci, the driver who plowed through a crowd of pedestrians, injuring one, last year at the annual ZombieWalk, held during Comic-Con International. He;s charged with felony reckless driving. Pocci, who's deaf, said he was frightened by the crowd, but prosecutors say he was angry and impatient. New video shows the car moving through the crowd and running over one woman. [NBC San Diego]
Political cartoons | J.J. Sedelmaier shows off some political cartoons by Winsor McCay on the topic of Prohibition, taken from a compilation, Temperance—or Prohibition?, that Sedelmaier picked up in a used bookstore. [Paste]
Political cartoons | On the 10th anniversary of the publication of cartoons of the Prophet Muhammad in the Danish newspaper Jyllands-Posten, experts and journalists reflect on the controversy that followed. While the publication of the cartoons and their aftermath have "made some journalists more thoughtful about the way they cover minority groups," journalism professor Angela Phillips says, "In other instances I think it made quite a lot of journalists if anything less sensitive to these issues. There is in some countries an even faster recourse to a rather essentialist notion of freedom of expression." [Agence France-Presse]
Creators | Masashi Kishimoto says he's done with Naruto and his friends, now that the manga has ended its 15-year run, and he's also not eager to return to the grind of a weekly series. However, that doesn't mean he's putting away his pen and ink. On the contrary, he has already created character designs for a new sci-fi series, and he's interested in working with the digital magazine Shonen Jump Plus, which would be more flexible with regard to the story's length and schedule. "Since Naruto was a bigger hit than I could ever imagine," he said, "I'd like to aim for the next hit. I don't know when I can announce the next manga, but because I plan on challenging myself to surpass Naruto, please wait for it." [Kotaku]
The Kramers Ergot anthology, edited by Sammy Harkham, has had an irregular publication history, with long spaces between volumes and a different publisher every few years. The oversized Kramers Ergot 7, which stood almost two feet tall, was released in 2008 by Buenaventura Press, and Kramers Ergot 8 came out in 2012 from the now-defunct art publisher PictureBox.
So it's big news that a new volume is on the way, this time from Fantagraphics.
Ed Piskor's bestselling book series Hip Hop Family Tree will released beginning in August as a monthly comic, a first for Fantagraphics Books. Each issue will feature a new cover and splash page, "director's commentary" from Piskor and other extras.
Debuting in 2013, the Eisner-nominated series chronicles the history of hip hop, tracing the genre back to its origins in the South Bronx. The 32-page first issue shines a spotlight on those break-dancers, graffiti artists, DJs and MCs who formed hip-hop culture in the early 1970s.
If you thought teaser images and announcements of announcements were strictly the domain of superhero publishers, think again.
Fantagraphics Books raised the subject of Daniel Clowes' next graphic novel with a teaser image posted on its Twitter page:
Passings | MAD Magazine writer Tom Koch passed away March 22 at age 89. He was a writer for the Bob and Ray radio comedy show in 1957, when MAD was trying to broaden its reach by featuring work by popular comedians. Koch adapted some routines he had written, and editor Al Feldstein realized his work was a good fit and asked him to to become a contribute. He wrote for the magazine for nearly four decades, contributing more than 300 pages, but he said he was proudest of a 1965 work, "43-man Squamish." It's still the magazine's most requested reprint. [News from ME, MAD Magazine]
At first glance, you might expect Lucy Knisley's latest travelogue Displacement to be filled with the same humor and insight as her previous books Relish: My Life in the Kitchen and An Age of License. After all, it recounts the 2012 cruise with her memory-impaired 91-year-old grandmother and physically challenged 93-year-old grandfather. Yet the quality that's made Knisley a great storyteller — her ability to recall nuanced encounters with a blend of wit and compassion — allows her to craft a compelling and complicated account of this time spent with her grandparents.
Auctions | A page of original artwork from 1971's Asterix and the Laurel Wreath sold at auction Sunday for more than $158,000, with proceeds going to benefit the families of those killed in the attack on Charlie Hebdo's offices. The art included a special dedication by Asterix co-creator Albert Uderzo, who came out of retirement in the days after the attack to draw tributes to the victims. The auction house Christie's waived its commission for Sunday's sale. [BBC News]
Political cartoons | Ecuadorean cartoonist Xavier Bonilla, who has been sued, threatened and reprimanded by his own government because of his political cartoons, revealed last week that he has also received threats from an Ecuadorean member of ISIS over a cartoon making fun of the extremist group. While he ultimately decided the threat wasn't credible, Bonilla said, "It has to be understood within this climate of hostility and harassment that's been created within the country. It's gotten to the point where even humor is being persecuted and oppressed by the president." Reporter Jim Wyss also looks at some other cases of government suppression of political cartoons in Latin America [Miami Herald]
As anyone who's ever worked at a newspaper can attest, readers don't react well to changes to the comics section, which is a major reason why so many strips trudge on, zombie-like, long after the spark of life left them. So when financial or space constraints force editors to eliminate some old favorites, they expect complaints — although not necessarily a profanity-laced tirade from an 8-year-old.
The Society of Illustrators has announced the winners of the 2015 Comic and Cartoon Art Annual competition. Olivier Schrauwen took the gold medal in the Long-Form category for Arsène Schrauwen, and Bianca Gagnarelli received top honors in the Short Form category for Fish. Lauren Weinstein won the gold medal in digital media for Carriers, her five-part webcomic about learning she and her husband both had the gene for cystic fibrosis, and therefore her unborn child might have the disease.
The winning entries will be put on display in two exhibits at the Society of Illustrators gallery in New York: The Short Form, Digital Media, and Special Format exhibit will run from June 16-July 18, and the Long Form, Single Image, and Comic Strip show will run from July 21-August 15. Many of the entries will also be on display at the MoCCA Arts Fest in April.
Here's the complete list of winners:
Following a complaint from a parent, officials at a New Mexico school district are investigating how a copy of Gilbert Hernandez's Palomar became part of a high school library's collection.
Catreena Lopez told KOAT Albuquerque she was disturbed by the "pornographic" images she found in the graphic novel, which her 14-year-old son reportedly checked out Wednesday from the Rio Rancho High School library, thinking it might be manga.
Flipping through the 500-page Fantagraphics hardcover, which collects all of Hernandez's inarguably mature-themed "Heartbreak Soup" stories from Love & Rockets, Lopez flagged 30 illustrations she considered to be pornographic.


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15 August 2006
Lord Turnbull, the judge who presided over Tommy Sheridan's defamation case against the News of the World, has formally complained to the Faculty of Advocates about Graeme Henderson, the advocate who initially represented Mr Sheridan.
Mr Henderson was sacked by Mr Sheridan after he questioned witness Anne Colvin about an alleged credit-card fraud for which he claimed she had served an 18-month jail term.
Ms Colvin, who had claimed she had seen Mr Sheridan, the former Scottish Socialist Party leader, taking part in group sex at a Glasgow hotel, said the fraud suggestion made by Mr Henderson was "ridiculous".
Mr Henderson then issued an apology, saying there was no grounds for his allegation. He was subsequently fired by Mr Sheridan.
Under the Faculty of Advocates' code of conduct an advocate must personally satisfy himself that there is evidence to support an allegation of criminal conduct before putting the allegation to a witness.
Ms Colvin's lawyer, Martha Rafferty of Glasgow firm Anthony Mahon, has separately lodged a formal complaint on Ms Colvin's behalf against Mr Henderson, and also against solicitors Bannatyne Kirkwood France & Co who then acted for Mr Sheridan.
The accusation made in court is protected by rules of privilege from any proceedings for defamation against Mr Henderson.
The Faculty's complaints committee has the power to order advocates to pay up to £5,000 in compensation to the complainer or suspend them from practice.
24 Jun 16
First Minister wants poll before article 50 Brexit time runs out
Undertaking to monitor negotiations closely and inform members
Holyrood ministers seek to maintain relationship with EU
23 Jun 16
Ray Macfarlane succeeds Iain Robertson
Delays in executry and litigation lead to compensation awards
Trial judge should have upheld no case to answer submission
22 Jun 16
Justices dismiss appeal as character of transfers "plain and obvious"
Response to SLC consultation calls for end to need for protective actions
Barrister elected to Strasbourg court by Council of Europe
Legal Wellbeing Scotland launches with 21 partners
21 Jun 16
Basic salaries 12% up in a year, though bonuses smaller
First such project in Scotland will provide specialist advice


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The ratings for the third Bellator show on Spike TV have been returned, and while the numbers are down slightly, the promotion still reached highs in one particularly strong category.
Industry sources confirmed to MMAWeekly.com that Bellator 87 averaged 705,000 viewers over the two hour broadcast on Thursday night.
The re-broadcast at 12 midnight pulled in additional 323,000 viewers, with the totals for both shows at 1.028 million viewers.
Now the 705,000 viewer mark is down from the first and second shows on Spike TV, but overall not a dramatic drop considering this card featured no title fights and also had a late change to the main event when lightweight Patricky "Pitbull" Freire was forced out of his scheduled bout.
One mark that bodes well for Bellator are the ratings in the coveted 18-34 male demographic where the reached a .6 rating, which is the highest for the promotion since moving to Spike TV three weeks ago.
Overall through 3 shows on Spike TV, Bellator has averages 858,000 viewers thus far.
Bobby Brents isn't looking past Shamrock FC opponent Bubba Grant, but he sees right through him with an eye on a Bellator contract. read news >>
Bubba Grant never gets too far ahead of himself, but winning a Shamrock FC belt and a Bellator contract could provide him with a breakout year. read news >>
Three contests have been added to Bellator 156: Galvao vs. Dantas 2 on June 17 at Save Mart Center in Fresno, California, completing the fight card. read news >>
Darrion Caldwell, a national college wrestling champion and one of the brightest up-and-coming bantamweights in MMA, will vie for a title shot this summer when he faces Joe Taimanglo in the main even read news >>
Bellator returns to Kansas with a pivotal matchup between undefeated phenom Darrion Caldwell and Joe Taimanglo to determine the No. 1 bantamweight contender. read news >>
Former Bellator MMA fighter Shahbulat Shamhalaev was hospitalized on Wednesday in Makhachkala, Russia after being shot "several times" while dining at a local restaurant. read news >>
Kevin "Kimbo Slice" Ferguson has passed away in a Florida hospital. Bellator MMA president Scott Coker confirmed the news on Monday evening. He was 42. read news >>
MMA fighter Kimbo Slice died Monday at the age of 42. Bellator President Scott Coker released a statement regarding the news, via B/R's Jonathan Snowden: TMZ Sports reported earlier Monday that polic read news >>
Arguably England's two top welterweights, Paul "Semtex" Daley (38-13-2) and Michael "Venom" Page (10-0) have each been assigned new opponents for the upcoming "Bellator 158: London" event. read news >>
An injury to Bellator 158 competitor Josh Koscheck has paved the way for a new co-main event at Bellator 156: Galvao vs. Dantas 2, pitting Chidi Njokuani against Thiago Jambo. read news >>


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Cubby O'Brien's Cab Toon Twirl, a favorite adventure of Sheriff Woody
The Cubby O'Brien's Cab Toon Twirl queue is like an adventure into the seedy side of Toontown. The adventure cabs are made to look like Beauty the Cab, and when you drive you can twirl them by hugging the tires.
Avoid Princess Mode
© copyright 2011–2016
A Pants Aflame production


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We're used to seeing smartphones with fun names come from the East, and while iBall's new handset may not have much under the hood, it is a winner in the nickname department. The iBall Andi 5N Dude is headed to India, and it will carry a price tag of Rs. 4,099 when it lands.
Samsung Galaxy Tab 4 Advanced gets benchmarked with mid-range specs
We don't see anyone knocking Apple from their throne as King of the Tablet world, but Samsung may eventually overcome them with their sheer numbers. A new tablet appears to be on the way called the Samsung Galaxy Tab 4 Advanced, and it has just passed through a benchmark listing with mediocre specs.
Humble Mobile Bundle 18 arrives with This War of Mine and Rebuild 3
We don't get to cover the Humble Bundles as much as we used to since the company decided to ditch their old release schedule. That said, we expected the offerings to improve and the Humble Mobile Bundle 18 is one that certainly will not disappoint.
Micromax Canvas Mega 2 specs and price at arrival
Many readers in India will have heard of the Micromax Canvas Mega that released last year. Now a successor to that smartphone has been launched in the form of the Mega 2. This is an affordable smartphone with a large 6-inch display, and you can find out about the Micromax Canvas Mega 2 specs, price […]
Meizu Pro 6 variant could be the Meizu Pro 6 Lite
The Meizu Pro 6 has been a smartphone we've covered plenty of times here at phonesreview.co.uk, but it appears the company isn't quite done with the handset yet. The Meizu Pro 6 Lite has mysteriously appeared online and looks to be a more affordable option to its sibling.
Moto X Pure Edition price now slashed by $100
Motorola's Moto X Pure Edition released last year as the US version of the high-end Moto X Style. We've already seen previous temporary price cuts for the device and now there's another good deal. The Moto X Pure Edition price has now been slashed by $100. However, if you want to take advantage you only […]
Lenovo Vibe S1 Marshmallow update arrives in India
The lengths of time it takes Android upgrades to reach different devices is a subject we often touch on at Phones Review. There's some good news today if you have Lenovo's Vibe S1 smartphone though, especially if you have been waiting for the Android 6.0 Marshmallow update. The Lenovo Vibe S1 Marshmallow update has now […]
HTC Desire 830 price made official at unveiling
It's only a week or so since we reported on leaked specs for the HTC Desire 830, and now the smartphone has just been officially launched in Taiwan. The HTC Desire 830 price has been made official at the unveiling, and we also have news on the confirmed specs, release date and more. There's always […]
Samsung Galaxy Tab E 8.0 on AT&T pre-order details
The Samsung Galaxy Tab E 8.0 was introduced early this year following on from the Galaxy Tab E 9.6 that released in 2015. Back in February we informed readers that the tablet has gone on sale at Verizon, and now we have news of it coming to another major carrier. Pre-orders for the Samsung Galaxy […]
VKWorld T1 6-inch smartphone tipped with $60 price
We seem to be seeing an increasing number of cheaper smartphones hit the market right now, with the majority coming out of China. Today we have some information about an upcoming phablet device with budget pricing. The VKWorld T1 6-inch smartphone is tipped with a $60 price, and that could make it a very appealing […]
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Fire union contract on City agenda
A proposed agreement between the labor union representing Clinton firefighters and the City of Clinton headlines the agenda for what appears to be a lengthy City Council meeting this evening (Tuesd
A proposed agreement between the labor union representing Clinton firefighters and the City of Clinton headlines the agenda for what appears to be a lengthy City Council meeting this evening (Tuesd
After a rocky start, it looks like the Arapaho-Butler Class of 2016 will get to go on their senior trip after all – even if it is a little later than they'd planned.
When parents worry about their children's safety nowadays, their top concerns are often things like how to keep them safe from online predators, along with teaching them to stay out of the street.
Area wheat farmers returned to the fields Wednesday and Thursday, trying to get the last of their 2016 crop in, as the price continued to drop.
Mackenzie Switzer, 7, helps her dad, Clinton firefighter Mark Switzer, with his helmet during a recent trip to the Clinton Fire Station.
Human remains found Sept. 6, 2015, have been identified as those of former Weatherford resident Nathan Sonobe, the Johnson County, Wyoming, Sheriff's Office confirmed Friday.
Clinton Daily News photographer Robert Bryan happened to be in the right place at the right time Thursday, when he and his camera captured a random act of kindness in progress.
A 2016-model Ford Taurus rented some time ago in Portland, Ore., was recovered here recently after sitting for three weeks on the parking lot of a Clinton motel.
Authorities are apparently still attempting to identify a skull found last week in Blaine County near Canton.
There were many eye-opening moments at the Frisco Center Tuesday as seniors learned just how low some people will go to scam them out of their money.
Plans for a new 10,000-
square-foot Dollar General store to be located in Clinton were announced Tuesday.
The death of a Clinton woman Tuesday has been determined to be a suicide, Sheriff's Investigator Kevin Evans said Wednesday.
After, by his own estimation, 41 years, one month and one week in the pulpit of the First Church of God, Roy Dobbs told his Church Council and congregation Sunday that he is retiring.
Two of the three co-valedictorians of the 2016 Clinton High School graduating class also were ato
522 Avant Avenue
Clinton, OK 73601
Phone: 580-323-5151
Fax: 580-323-5154


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Houses to let in Wolton Road, Kesgrave, Ipswich IP5
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Marshall Close, Kesgrave, Ipswich IP5End terrace house to rent - 2 bedrooms
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Austwick berry are pleased to offer to let this attractive 2 bedroom end of terrace home. The property is situated down the end of a quiet cul de sac on the highly regarded grange farm development. The property is in very good condition. Internally ...
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The Lloyds, Kesgrave IP5Link-detached house to rent - 3 bedrooms
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Unfurnished three bedroom detached family home situated in a quiet cul-de-sac position in the very popular area of Grange farm, Kesgrave. A small pet maybe considered with an extra pet deposit of £250. Housing benefit is not accepted.
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Release Date:
September 1, 2001
Date Published:
September 1, 2001
What happens when the five luckiest children in the entire world walk through the doors of Willy Wonka's famous, mysterious chocolate factory? What happens when, one by one, the children disobey Mr. Wonka's orders? In Dahl's most popular story, the nasty are punished and the good are deliciously, sumptuously rewarded.
Darkly humorous, slightly creepy, and ultimately the tale of a nice guy who just might finish first, CHARLIE AND THE CHOCOLATE FACTORY is one of the prolific Roald Dahl's most popular works. Life changes for a poor but kindhearted boy named Charlie Bucket when he manages to win one of five golden tickets that allows him, and a guest of his choice to tour the chocolate factory of an eccentric and reclusive candy inventor named Willy Wonka. On the big day Charlie and the other visitors are warned to obey all of Mr. Wonka's strict, but rather strange, rules for behavior. What will happen if they disobey? And why has Mr. Wonka allowed them to tour his factory in the first place? Illustrated with B&W drawings, CHARLIE AND THE CHOCOLATE FACTORY introduces readers to such unforgettable characters as the hardworking Oompa-Loompas, the extremely spoiled Veruca Salt, and, as readers will soon discover, the aptly named Violet Beauregarde.
Genre: Juvenile Nonfiction, Juvenile Fiction
Sub-Genre: Science Fiction + Fantasy + Magic, Humorous Stories
- Genre: Juvenile Fiction, Juvenile Nonfiction
- Subgenre: Humorous Stories, Science Fiction + Fantasy + Magic
- Age: 9-12 years
- Publisher: Alfred a Knopf Inc
- Edition: Revised
- Language: English
- Release Date: September 1, 2001
- Author: Roald Dahl
- Illustrator: Quentin Blake
other Info.
- Online Item #: 11960666
- Store Item Number (DPCI): 247-03-0965
- Item can be gift wrapped.
- Made in the USA or Imported


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2013 Web Spotlights
G3: Genes|Genomes|Genetics publishes high-quality, valuable findings, regardless of perceived impact. G3 publishes foundational research that generates useful genetic and genomic information such as genome maps, single gene studies, QTL studies, mutant screens and advances in methods and technology, novel mutant collections, genome-wide association studies (GWAS) including gene expression, SNP, and CNV studies; exome sequences related to a specific disease but lacking functional follow-up, personal exome and genome sequencing case, disease, and population reports, and more.
Conceived by the Genetics Society of America, with its first issue published June 2011, G3 is fully open access. G3 uses a Creative Commons license that allows the most free use of the data, which anyone can download, analyze, mine, and reuse, provided that the authors of the article receive credit. GSA believes that rapid dissemination of useful data is the necessary foundation for analysis that leads to mechanistic insights. It is our hope is that this strategy will spawn new discovery.
Like GENETICS, G3 is fast—with a 31-day turnaround time from submission to first decision—and rapid time-to-publication. And like GENETICS, G3 manuscripts are thoroughly peer-reviewed, with careful decisions made by practicing scientists. Before publication, G3 articles receive a thorough copy-edit, ensuring that articles enjoy maximum clarity and impact.
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Wednesday, June 1 2016 08:52:57 AM
Sdt97: A Point Mutation in the 5' Untranslated Region Confers Semidwarfism in Rice
Tong, J., Han, Z., Han, A., Liu, X., Zhang, S., Fu, B., Hu, J., Su, J., Li, S., Wang, S., Zhu, Y.
Semidwarfism is an important agronomic trait in rice breeding programs. The semidwarf mutant gene Sdt97 was previously described. However, the molecular mechanism underlying the mutant is yet to be elucidated. In this study, we identified the mutant gene by a map-based cloning method. Using a residual heterozygous line (RHL) population, Sdt97 was mapped to the long arm of chromosome 6 in the interval of nearly 60 kb between STS marker N6 and SNP marker N16 within the PAC clone P0453H04. Sequencing of the candidate genes in the target region revealed that a base transversion from G to C occurred in the 5' untranslated region of Sdt97. qRT-PCR results confirmed that the transversion induced an obvious change in the expression pattern of Sdt97 at different growth and developmental stages. Plants transgenic for Sdt97 resulted in the restoration of semidwarfism of the mutant phenotype, or displayed a greater dwarf phenotype than the mutant. Our results indicate that a point mutation in the 5' untranslated region of Sdt97 confers semidwarfism in rice. Functional analysis of Sdt97 will open a new field of study for rice semidwarfism, and also expand our knowledge of the molecular mechanism of semidwarfism in rice.
Several Critical Cell Types, Tissues, and Pathways Are Implicated in Genome-Wide Association Studies for Systemic Lupus Erythematosus
Liu, L., Yin, X., Wen, L., Yang, C., Sheng, Y., Lin, Y., Zhu, Z., Shen, C., Shi, Y., Zheng, Y., Yang, S., Zhang, X., Cui, Y.
We aimed to elucidate the cell types, tissues, and pathways influenced by common variants in systemic lupus erythematosus (SLE). We applied a nonparameter enrichment statistical approach, termed SNPsea, in 181 single nucleotide polymorphisms (SNPs) that have been identified to be associated with the risk of SLE through genome-wide association studies (GWAS) in Eastern Asian and Caucasian populations, to manipulate the critical cell types, tissues, and pathways. In the two most significant cells' findings (B lymphocytes and CD14+ monocytes), we subjected the GWAS association evidence in the Han Chinese population to an enrichment test of expression quantitative trait locus (QTL) sites and DNase I hypersensitivity, respectively. In both Eastern Asian and Caucasian populations, we observed that the expression level of SLE GWAS implicated genes was significantly elevated in xeroderma pigentosum B cells (P ≤ 1.00 10–6), CD14+ monocytes (P ≤ 2.74 10–4) and CD19+ B cells (P ≤ 2.00 10–6), and plasmacytoid dendritic cells (pDCs) (P ≤ 9.00 10–6). We revealed that the SLE GWAS-associated variants were more likely to reside in expression QTL in B lymphocytes (q1/q0 = 2.15, P = 1.23 10–44) and DNase I hypersensitivity sites (DHSs) in CD14+ monocytes (q1/q0 = 1.41, P = 0.08). We observed the common variants affected the risk of SLE mostly through by regulating multiple immune system processes and immune response signaling. This study sheds light on several immune cells and responses, as well as the regulatory effect of common variants in the pathogenesis of SLE.
Interconnections Between RNA-Processing Pathways Revealed by a Sequencing-Based Genetic Screen for Pre-mRNA Splicing Mutants in Fission Yeast
Larson, A., Fair, B. J., Pleiss, J. A.
Pre-mRNA splicing is an essential component of eukaryotic gene expression and is highly conserved from unicellular yeasts to humans. Here, we present the development and implementation of a sequencing-based reverse genetic screen designed to identify nonessential genes that impact pre-mRNA splicing in the fission yeast Schizosaccharomyces pombe, an organism that shares many of the complex features of splicing in higher eukaryotes. Using a custom-designed barcoding scheme, we simultaneously queried ~3000 mutant strains for their impact on the splicing efficiency of two endogenous pre-mRNAs. A total of 61 nonessential genes were identified whose deletions resulted in defects in pre-mRNA splicing; enriched among these were factors encoding known or predicted components of the spliceosome. Included among the candidates identified here are genes with well-characterized roles in other RNA-processing pathways, including heterochromatic silencing and 3' end processing. Splicing-sensitive microarrays confirm broad splicing defects for many of these factors, revealing novel functional connections between these pathways.
Local Ancestry Inference in a Large US-Based Hispanic/Latino Study: Hispanic Community Health Study/Study of Latinos (HCHS/SOL)
Browning, S. R., Grinde, K., Plantinga, A., Gogarten, S. M., Stilp, A. M., Kaplan, R. C., Aviles-Santa, M. L., Browning, B. L., Laurie, C. C.
We estimated local ancestry on the autosomes and X chromosome in a large US-based study of 12,793 Hispanic/Latino individuals using the RFMix method, and we compared different reference panels and approaches to local ancestry estimation on the X chromosome by means of Mendelian inconsistency rates as a proxy for accuracy. We developed a novel and straightforward approach to performing ancestry-specific PCA after finding artifactual behavior in the results from an existing approach. Using the ancestry-specific PCA, we found significant population structure within African, European, and Amerindian ancestries in the Hispanic/Latino individuals in our study. In the African ancestral component of the admixed individuals, individuals whose grandparents were from Central America clustered separately from individuals whose grandparents were from the Caribbean, and also from reference Yoruba and Mandenka West African individuals. In the European component, individuals whose grandparents were from Puerto Rico diverged partially from other background groups. In the Amerindian ancestral component, individuals clustered into multiple different groups depending on the grandparental country of origin. Therefore, local ancestry estimation provides further insight into the complex genetic structure of US Hispanic/Latino populations, which must be properly accounted for in genotype-phenotype association studies. It also provides a basis for admixture mapping and ancestry-specific allele frequency estimation, which are useful in the identification of risk factors for disease.
Genomic Signatures of Experimental Adaptation to Antimicrobial Peptides in Staphylococcus aureus
Johnston, P. R., Dobson, A. J., Rolff, J.
The evolution of resistance against antimicrobial peptides has long been considered unlikely due to their mechanism of action, yet experimental selection with antimicrobial peptides (AMPs) results in rapid evolution of resistance in several species of bacteria. Although numerous studies have utilized mutant screens to identify loci that determine AMP susceptibility, there is a dearth of data concerning the genomic changes that accompany experimental evolution of AMP resistance. Using genome resequencing, we analyzed the mutations that arose during experimental evolution of resistance to the cationic AMPs iseganan, melittin, and pexiganan, as well as to a combination of melittin and pexiganan, or to the aminoglycoside antibiotic streptomycin. Analysis of 17 independently replicated Staphylococcus aureus selection lines, including unselected controls, showed that each AMP selected for mutations at distinct loci. We identify mutations in genes involved in the synthesis and maintenance of the cell envelope. These include genes previously identified from mutant screens for AMP resistance, and genes involved in the response to AMPs and cell-wall-active antibiotics. Furthermore, transposon insertion mutants were used to verify that a number of the identified genes are directly involved in determining AMP susceptibility. Strains selected for AMP resistance under controlled experimental evolution displayed consistent AMP-specific mutations in genes that determine AMP susceptibility. This suggests that different routes to evolve resistance are favored within a controlled genetic background.
Identification and Characterization of a cis-Regulatory Element for Zygotic Gene Expression in Chlamydomonas reinhardtii
Hamaji, T., Lopez, D., Pellegrini, M., Umen, J.
Upon fertilization Chlamydomonas reinhardtii zygotes undergo a program of differentiation into a diploid zygospore that is accompanied by transcription of hundreds of zygote-specific genes. We identified a distinct sequence motif we term a zygotic response element (ZYRE) that is highly enriched in promoter regions of C. reinhardtii early zygotic genes. A luciferase reporter assay was used to show that native ZYRE motifs within the promoter of zygotic gene ZYS3 or intron of zygotic gene DMT4 are necessary for zygotic induction. A synthetic luciferase reporter with a minimal promoter was used to show that ZYRE motifs introduced upstream are sufficient to confer zygotic upregulation, and that ZYRE-controlled zygotic transcription is dependent on the homeodomain transcription factor GSP1. We predict that ZYRE motifs will correspond to binding sites for the homeodomain proteins GSP1-GSM1 that heterodimerize and activate zygotic gene expression in early zygotes.
Quantitative Trait Locus Analysis of Mating Behavior and Male Sex Pheromones in Nasonia Wasps
Diao, W., Mousset, M., Horsburgh, G. J., Vermeulen, C. J., Johannes, F., van de Zande, L., Ritchie, M. G., Schmitt, T., Beukeboom, L. W.
A major focus in speciation genetics is to identify the chromosomal regions and genes that reduce hybridization and gene flow. We investigated the genetic architecture of mating behavior in the parasitoid wasp species pair Nasonia giraulti and Nasonia oneida that exhibit strong prezygotic isolation. Behavioral analysis showed that N. oneida females had consistently higher latency times, and broke off the mating sequence more often in the mounting stage when confronted with N. giraulti males compared with males of their own species. N. oneida males produce a lower quantity of the long-range male sex pheromone (4R,5S)-5-hydroxy-4-decanolide (RS-HDL). Crosses between the two species yielded hybrid males with various pheromone quantities, and these males were used in mating trials with females of either species to measure female mate discrimination rates. A quantitative trait locus (QTL) analysis involving 475 recombinant hybrid males (F2), 2148 reciprocally backcrossed females (F3), and a linkage map of 52 equally spaced neutral single nucleotide polymorphism (SNP) markers plus SNPs in 40 candidate mating behavior genes revealed four QTL for male pheromone amount, depending on partner species. Our results demonstrate that the RS-HDL pheromone plays a role in the mating system of N. giraulti and N. oneida, but also that additional communication cues are involved in mate choice. No QTL were found for female mate discrimination, which points at a polygenic architecture of female choice with strong environmental influences.
New Software for the Fast Estimation of Population Recombination Rates (FastEPRR) in the Genomic Era
Gao, F., Ming, C., Hu, W., Li, H.
Genetic recombination is a very important evolutionary mechanism that mixes parental haplotypes and produces new raw material for organismal evolution. As a result, information on recombination rates is critical for biological research. In this paper, we introduce a new extremely fast open-source software package (FastEPRR) that uses machine learning to estimate recombination rate $$\rho $$ (=$$4{N}_{e}r$$) from intraspecific DNA polymorphism data. When $$\rho > 10$$ and the number of sampled diploid individuals is large enough ($$\ge 50$$), the variance of $${\rho }_{\hbox{ FastEPRR }}$$ remains slightly smaller than that of $${\rho }_{\hbox{ LDhat }}$$. The new estimate $${\rho }_{\hbox{ comb }}$$ (calculated by averaging $${\rho }_{\hbox{ FastEPRR }}$$ and $${\rho }_{\hbox{ LDhat }}$$) has the smallest variance of all cases. When estimating $${\rho }_{\hbox{ FastEPRR }}$$, the finite-site model was employed to analyze cases with a high rate of recurrent mutations, and an additional method is proposed to consider the effect of variable recombination rates within windows. Simulations encompassing a wide range of parameters demonstrate that different evolutionary factors, such as demography and selection, may not increase the false positive rate of recombination hotspots. Overall, accuracy of FastEPRR is similar to the well-known method, LDhat, but requires far less computation time. Genetic maps for each human population (YRI, CEU, and CHB) extracted from the 1000 Genomes OMNI data set were obtained in less than 3 d using just a single CPU core. The Pearson Pairwise correlation coefficient between the $${\rho }_{\hbox{ FastEPRR }}$$ and $${\rho }_{\hbox{ LDhat }}$$ maps is very high, ranging between 0.929 and 0.987 at a 5-Mb scale. Considering that sample sizes for these kinds of data are increasing dramatically with advances in next-generation sequencing technologies, FastEPRR (freely available at http://www.picb.ac.cn/evolgen/) is expected to become a widely used tool for establishing genetic maps and studying recombination hotspots in the population genomic era.
Patterns of Genome-Wide Variation in Glossina fuscipes fuscipes Tsetse Flies from Uganda
Gloria-Soria, A., Dunn, W. A., Telleria, E. L., Evans, B. R., Okedi, L., Echodu, R., Warren, W. C., Montague, M. J., Aksoy, S., Caccone, A.
The tsetse fly Glossina fuscipes fuscipes (Gff) is the insect vector of the two forms of Human African Trypanosomiasis (HAT) that exist in Uganda. Understanding Gff population dynamics, and the underlying genetics of epidemiologically relevant phenotypes is key to reducing disease transmission. Using ddRAD sequence technology, complemented with whole-genome sequencing, we developed a panel of ~73,000 single-nucleotide polymorphisms (SNPs) distributed across the Gff genome that can be used for population genomics and to perform genome-wide-association studies. We used these markers to estimate genomic patterns of linkage disequilibrium (LD) in Gff, and used the information, in combination with outlier-locus detection tests, to identify candidate regions of the genome under selection. LD in individual populations decays to half of its maximum value (r2max/2) between 1359 and 2429 bp. The overall LD estimated for the species reaches r2max/2 at 708 bp, an order of magnitude slower than in Drosophila. Using 53 infected (Trypanosoma spp.) and uninfected flies from four genetically distinct Ugandan populations adapted to different environmental conditions, we were able to identify SNPs associated with the infection status of the fly and local environmental adaptation. The extent of LD in Gff likely facilitated the detection of loci under selection, despite the small sample size. Furthermore, it is probable that LD in the regions identified is much higher than the average genomic LD due to strong selection. Our results show that even modest sample sizes can reveal significant genetic associations in this species, which has implications for future studies given the difficulties of collecting field specimens with contrasting phenotypes for association analysis.
Fungal Innate Immunity Induced by Bacterial Microbe-Associated Molecular Patterns (MAMPs)
Ipcho, S., Sundelin, T., Erbs, G., Kistler, H. C., Newman, M.-A., Olsson, S.
Plants and animals detect bacterial presence through Microbe-Associated Molecular Patterns (MAMPs) which induce an innate immune response. The field of fungal–bacterial interaction at the molecular level is still in its infancy and little is known about MAMPs and their detection by fungi. Exposing Fusarium graminearum to bacterial MAMPs led to increased fungal membrane hyperpolarization, a putative defense response, and a range of transcriptional responses. The fungus reacted with a different transcript profile to each of the three tested MAMPs, although a core set of genes related to energy generation, transport, amino acid production, secondary metabolism, and especially iron uptake were detected for all three. Half of the genes related to iron uptake were predicted MirA type transporters that potentially take up bacterial siderophores. These quick responses can be viewed as a preparation for further interactions with beneficial or pathogenic bacteria, and constitute a fungal innate immune response with similarities to those of plants and animals.
Comparative Genomics of Interreplichore Translocations in Bacteria: A Measure of Chromosome Topology?
Khedkar, S., Seshasayee, A. S. N.
Genomes evolve not only in base sequence but also in terms of their architecture, defined by gene organization and chromosome topology. Whereas genome sequence data inform us about the changes in base sequences for a large variety of organisms, the study of chromosome topology is restricted to a few model organisms studied using microscopy and chromosome conformation capture techniques. Here, we exploit whole genome sequence data to study the link between gene organization and chromosome topology in bacteria. Using comparative genomics across ~250 pairs of closely related bacteria we show that: (a) many organisms show a high degree of interreplichore translocations throughout the chromosome and not limited to the inversion-prone terminus (ter) or the origin of replication (oriC); (b) translocation maps may reflect chromosome topologies; and (c) symmetric interreplichore translocations do not disrupt the distance of a gene from oriC or affect gene expression states or strand biases in gene densities. In summary, we suggest that translocation maps might be a first line in defining a gross chromosome topology given a pair of closely related genome sequences.
A High-Resolution SNP Array-Based Linkage Map Anchors a New Domestic Cat Draft Genome Assembly and Provides Detailed Patterns of Recombination
Li, G., Hillier, L. W., Grahn, R. A., Zimin, A. V., David, V. A., Menotti-Raymond, M., Middleton, R., Hannah, S., Hendrickson, S., Makunin, A., OBrien, S. J., Minx, P., Wilson, R. K., Lyons, L. A., Warren, W. C., Murphy, W. J.
High-resolution genetic and physical maps are invaluable tools for building accurate genome assemblies, and interpreting results of genome-wide association studies (GWAS). Previous genetic and physical maps anchored good quality draft assemblies of the domestic cat genome, enabling the discovery of numerous genes underlying hereditary disease and phenotypes of interest to the biomedical science and breeding communities. However, these maps lacked sufficient marker density to order thousands of shorter scaffolds in earlier assemblies, which instead relied heavily on comparative mapping with related species. A high-resolution map would aid in validating and ordering chromosome scaffolds from existing and new genome assemblies. Here, we describe a high-resolution genetic linkage map of the domestic cat genome based on genotyping 453 domestic cats from several multi-generational pedigrees on the Illumina 63K SNP array. The final maps include 58,055 SNP markers placed relative to 6637 markers with unique positions, distributed across all autosomes and the X chromosome. Our final sex-averaged maps span a total autosomal length of 4464 cM, the longest described linkage map for any mammal, confirming length estimates from a previous microsatellite-based map. The linkage map was used to order and orient the scaffolds from a substantially more contiguous domestic cat genome assembly (Felis catus v8.0), which incorporated ~20 x coverage of Illumina fragment reads. The new genome assembly shows substantial improvements in contiguity, with a nearly fourfold increase in N50 scaffold size to 18 Mb. We use this map to report probable structural errors in previous maps and assemblies, and to describe features of the recombination landscape, including a massive (~50 Mb) recombination desert (of virtually zero recombination) on the X chromosome that parallels a similar desert on the porcine X chromosome in both size and physical location.
mRNA-Associated Processes and Their Influence on Exon-Intron Structure in Drosophila melanogaster
Lepennetier, G., Catania, F.
mRNA-associated processes and gene structure in eukaryotes are typically treated as separate research subjects. Here, we bridge this separation and leverage the extensive multidisciplinary work on Drosophila melanogaster to examine the roles that capping, splicing, cleavage/polyadenylation, and telescripting (i.e., the protection of nascent transcripts from premature cleavage/polyadenylation by the splicing factor U1) might play in shaping exon-intron architecture in protein-coding genes. Our findings suggest that the distance between subsequent internal 5' splice sites (5'ss) in Drosophila genes is constrained such that telescripting effects are maximized, in theory, and thus nascent transcripts are less vulnerable to premature termination. Exceptionally weak 5'ss and constraints on intron-exon size at the gene 5' end also indicate that capping might enhance the recruitment of U1 and, in turn, promote telescripting at this location. Finally, a positive correlation between last exon length and last 5'ss strength suggests that optimal donor splice sites in the proximity of the pre-mRNA tail may inhibit the processing of downstream polyadenylation signals more than weak donor splice sites do. These findings corroborate and build upon previous experimental and computational studies on Drosophila genes. They support the possibility, hitherto scantly explored, that mRNA-associated processes impose significant constraints on the evolution of eukaryotic gene structure.
The Immature Fiber Mutant Phenotype of Cotton (Gossypium hirsutum) Is Linked to a 22-bp Frame-Shift Deletion in a Mitochondria Targeted Pentatricopeptide Repeat Gene
Thyssen, G. N., Fang, D. D., Zeng, L., Song, X., Delhom, C. D., Condon, T. L., Li, P., Kim, H. J.
Cotton seed trichomes are the most important source of natural fibers globally. The major fiber thickness properties influence the price of the raw material, and the quality of the finished product. The recessive immature fiber (im) gene reduces the degree of fiber cell wall thickening by a process that was previously shown to involve mitochondrial function in allotetraploid Gossypium hirsutum. Here, we present the fine genetic mapping of the im locus, gene expression analysis of annotated proteins near the locus, and association analysis of the linked markers. Mapping-by-sequencing identified a 22-bp deletion in a pentatricopeptide repeat (PPR) gene that is completely linked to the immature fiber phenotype in 2837 F2 plants, and is absent from all 163 cultivated varieties tested, although other closely linked marker polymorphisms are prevalent in the diversity panel. This frame-shift mutation results in a transcript with two long open reading frames: one containing the N-terminal transit peptide that targets mitochondria, the other containing only the RNA-binding PPR domains, suggesting that a functional PPR protein cannot be targeted to mitochondria in the im mutant. Taken together, these results suggest that PPR gene Gh_A03G0489 is involved in the cotton fiber wall thickening process, and is a promising candidate gene at the im locus. Our findings expand our understanding of the molecular mechanisms that modulate cotton fiber fineness and maturity, and may facilitate the development of cotton varieties with superior fiber attributes.
Multi-Population Selective Genotyping to Identify Soybean [Glycine max (L.) Merr.] Seed Protein and Oil QTLs
Phansak, P., Soonsuwon, W., Hyten, D. L., Song, Q., Cregan, P. B., Graef, G. L., Specht, J. E.
Plant breeders continually generate ever-higher yielding cultivars, but also want to improve seed constituent value, which is mainly protein and oil, in soybean [Glycine max (L.) Merr.]. Identification of genetic loci governing those two traits would facilitate that effort. Though genome-wide association offers one such approach, selective genotyping of multiple biparental populations offers a complementary alternative, and was evaluated here, using 48 F2:3 populations (n = ~224 plants) created by mating 48 high protein germplasm accessions to cultivars of similar maturity, but with normal seed protein content. All F2:3 progeny were phenotyped for seed protein and oil, but only 22 high and 22 low extreme progeny in each F2:3 phenotypic distribution were genotyped with a 1536-SNP chip (ca. 450 bimorphic SNPs detected per mating). A significant quantitative trait locus (QTL) on one or more chromosomes was detected for protein in 35 (73%), and for oil in 25 (52%), of the 48 matings, and these QTL exhibited additive effects of ≥ 4 g kg–1 and R2 values of 0.07 or more. These results demonstrated that a multiple-population selective genotyping strategy, when focused on matings between parental phenotype extremes, can be used successfully to identify germplasm accessions possessing large-effect QTL alleles. Such accessions would be of interest to breeders to serve as parental donors of those alleles in cultivar development programs, though 17 of the 48 accessions were not unique in terms of SNP genotype, indicating that diversity among high protein accessions in the germplasm collection is less than what might ordinarily be assumed.
Saccharomyces cerevisiae Tti2 Regulates PIKK Proteins and Stress Response
Hoffman, K. S., Duennwald, M. L., Karagiannis, J., Genereaux, J., McCarton, A. S., Brandl, C. J.
The TTT complex is composed of the three essential proteins Tel2, Tti1, and Tti2. The complex is required to maintain steady state levels of phosphatidylinositol 3-kinase-related kinase (PIKK) proteins, including mTOR, ATM/Tel1, ATR/Mec1, and TRRAP/Tra1, all of which serve as regulators of critical cell signaling pathways. Due to their association with heat shock proteins, and with newly synthesized PIKK peptides, components of the TTT complex may act as cochaperones. Here, we analyze the consequences of depleting the cellular level of Tti2 in Saccharomyces cerevisiae. We show that yeast expressing low levels of Tti2 are viable under optimal growth conditions, but the cells are sensitive to a number of stress conditions that involve PIKK pathways. In agreement with this, depleting Tti2 levels decreased expression of Tra1, Mec1, and Tor1, affected their localization and inhibited the stress responses in which these molecules are involved. Tti2 expression was not increased during heat shock, implying that it does not play a general role in the heat shock response. However, steady state levels of Hsp42 increase when Tti2 is depleted, and tti2L187P has a synthetic interaction with exon 1 of the human Huntingtin gene containing a 103 residue polyQ sequence, suggesting a general role in protein quality control. We also find that overexpressing Hsp90 or its cochaperones is synthetic lethal when Tti2 is depleted, an effect possibly due to imbalanced stoichiometry of a complex required for PIKK assembly. These results indicate that Tti2 does not act as a general chaperone, but may have a specialized function in PIKK folding and/or complex assembly.
The Histone Variant H3.3 Is Enriched at Drosophila Amplicon Origins but Does Not Mark Them for Activation
Paranjape, N. P., Calvi, B. R.
Eukaryotic DNA replication begins from multiple origins. The origin recognition complex (ORC) binds origin DNA and scaffolds assembly of a prereplicative complex (pre-RC), which is subsequently activated to initiate DNA replication. In multicellular eukaryotes, origins do not share a strict DNA consensus sequence, and their activity changes in concert with chromatin status during development, but mechanisms are ill-defined. Previous genome-wide analyses in Drosophila and other organisms have revealed a correlation between ORC binding sites and the histone variant H3.3. This correlation suggests that H3.3 may designate origin sites, but this idea has remained untested. To address this question, we examined the enrichment and function of H3.3 at the origins responsible for developmental gene amplification in the somatic follicle cells of the Drosophila ovary. We found that H3.3 is abundant at these amplicon origins. H3.3 levels remained high when replication initiation was blocked, indicating that H3.3 is abundant at the origins before activation of the pre-RC. H3.3 was also enriched at the origins during early oogenesis, raising the possibility that H3.3 bookmarks sites for later amplification. However, flies null mutant for both of the H3.3 genes in Drosophila did not have overt defects in developmental gene amplification or genomic replication, suggesting that H3.3 is not essential for the assembly or activation of the pre-RC at origins. Instead, our results imply that the correlation between H3.3 and ORC sites reflects other chromatin attributes that are important for origin function.
A Genetic Map Between Gossypium hirsutum and the Brazilian Endemic G. mustelinum and Its Application to QTL Mapping
Wang, B., Liu, L., Zhang, D., Zhuang, Z., Guo, H., Qiao, X., Wei, L., Rong, J., May, O. L., Paterson, A. H., Chee, P. W.
Among the seven tetraploid cotton species, little is known about transmission genetics and genome organization in Gossypium mustelinum, the species most distant from the source of most cultivated cotton, G. hirsutum. In this research, an F2 population was developed from an interspecific cross between G. hirsutum and G. mustelinum (HM). A genetic linkage map was constructed mainly using simple sequence repeat (SSRs) and restriction fragment length polymorphism (RFLP) DNA markers. The arrangements of most genetic loci along the HM chromosomes were identical to those of other tetraploid cotton species. However, both major and minor structural rearrangements were also observed, for which we propose a parsimony-based model for structural divergence of tetraploid cottons from common ancestors. Sequences of mapped markers were used for alignment with the 26 scaffolds of the G. hirsutum draft genome, and showed high consistency. Quantitative trait locus (QTL) mapping of fiber elongation in advanced backcross populations derived from the same parents demonstrated the value of the HM map. The HM map will serve as a valuable resource for QTL mapping and introgression of G. mustelinum alleles into G. hirsutum, and help clarify evolutionary relationships between the tetraploid cotton genomes.
The Drosophila prage Gene, Required for Maternal Transcript Destabilization in Embryos, Encodes a Predicted RNA Exonuclease
Cui, J., Lai, Y. W., Sartain, C. V., Zuckerman, R. M., Wolfner, M. F.
Egg activation, the transition of mature oocytes into developing embryos, is critical for the initiation of embryogenesis. This process is characterized by resumption of meiosis, changes in the egg's coverings and by alterations in the transcriptome and proteome of the egg; all of these occur in the absence of new transcription. Activation of the egg is prompted by ionic changes in the cytoplasm (usually a rise in cytosolic calcium levels) that are triggered by fertilization in some animals and by mechanosensitive cues in others. The egg's transcriptome is dramatically altered during the process, including by the removal of many maternal mRNAs that are not needed for embryogenesis. However, the mechanisms and regulators of this selective RNA degradation are not yet fully known. Forward genetic approaches in Drosophila have identified maternal-effect genes whose mutations prevent the transcriptome changes. One of these genes, prage (prg), was identified by Tadros et al. in a screen for mutants that fail to destabilize maternal transcripts. We identified the molecular nature of the prg gene through a combination of deficiency mapping, complementation analysis, and DNA sequencing of both extant prg mutant alleles. We find that prg encodes a ubiquitously expressed predicted exonuclease, consistent with its role in maternal mRNA destabilization during egg activation.
Synthetic Ligands of Cannabinoid Receptors Affect Dauer Formation in the Nematode Caenorhabditis elegans
Reis Rodrigues, P., Kaul, T. K., Ho, J.-H., Lucanic, M., Burkewitz, K., Mair, W. B., Held, J. M., Bohn, L. M., Gill, M. S.
Under adverse environmental conditions the nematode Caenorhabditis elegans can enter an alternate developmental stage called the dauer larva. To identify lipophilic signaling molecules that influence this process, we screened a library of bioactive lipids and found that AM251, an antagonist of the human cannabinoid (CB) receptor, suppresses dauer entry in daf-2 insulin receptor mutants. AM251 acted synergistically with glucose supplementation indicating that the metabolic status of the animal influenced the activity of this compound. Similarly, loss of function mutations in the energy-sensing AMP-activated kinase subunit, aak-2, enhanced the dauer-suppressing effects of AM251, while constitutive activation of aak-2 in neurons was sufficient to inhibit AM251 activity. Chemical epistasis experiments indicated that AM251 acts via G-protein signaling and requires the TGF-β ligand DAF-7, the insulin peptides DAF-28 and INS-6, and a functional ASI neuron to promote reproductive growth. AM251 also required the presence of the SER-5 serotonin receptor, but in vitro experiments suggest that this may not be via a direct interaction. Interestingly, we found that other antagonists of mammalian CB receptors also suppress dauer entry, while the nonselective CB receptor agonist, O-2545, not only inhibited the activity of AM251, but also was able to promote dauer entry when administered alone. Since worms do not have obvious orthologs of CB receptors, the effects of synthetic CBs on neuroendocrine signaling in C. elegans are likely to be mediated via another, as yet unknown, receptor mechanism. However, we cannot exclude the existence of a noncanonical CB receptor in C. elegans.
Evaluation of IRX Genes and Conserved Noncoding Elements in a Region on 5p13.3 Linked to Families with Familial Idiopathic Scoliosis and Kyphosis
Justice, C. M., Bishop, K., Carrington, B., Mullikin, J. C., Swindle, K., Marosy, B., Sood, R., Miller, N. H., Wilson, A. F.
Because of genetic heterogeneity present in idiopathic scoliosis, we previously defined clinical subsets (a priori) from a sample of families with idiopathic scoliosis to find genes involved with spinal curvature. Previous genome-wide linkage analysis of seven families with at least two individuals with kyphoscoliosis found linkage (P-value = 0.002) in a 3.5-Mb region on 5p13.3 containing only three known genes, IRX1, IRX2, and IRX4. In this study, the exons of IRX1, IRX2, and IRX4, the conserved noncoding elements in the region, and the exons of a nonprotein coding RNA, LOC285577, were sequenced. No functional sequence variants were identified. An intrafamilial test of association found several associated noncoding single nucleotide variants. The strongest association was with rs12517904 (P = 0.00004), located 6.5 kb downstream from IRX1. In one family, the genotypes of nine variants differed from the reference allele in all individuals with kyphoscoliosis, and two of three individuals with scoliosis, but did not differ from the reference allele in all other genotyped individuals. One of these variants, rs117273909, was located in a conserved noncoding region that functions as an enhancer in mice. To test whether the variant allele at rs117273909 had an effect on enhancer activity, zebrafish transgenesis was performed with overlapping fragments of 198 and 687 bp containing either the wild type or the variant allele. Our data suggests that this region acts as a regulatory element; however, its size and target gene(s) need to be identified to determine its role in idiopathic scoliosis.
Genetic Interactions Between the Meiosis-Specific Cohesin Components, STAG3, REC8, and RAD21L
Ward, A., Hopkins, J., Mckay, M., Murray, S., Jordan, P. W.
Cohesin is an essential structural component of chromosomes that ensures accurate chromosome segregation during mitosis and meiosis. Previous studies have shown that there are cohesin complexes specific to meiosis, required to mediate homologous chromosome pairing, synapsis, recombination, and segregation. Meiosis-specific cohesin complexes consist of two structural maintenance of chromosomes proteins (SMC1α/SMC1β and SMC3), an α-kleisin protein (RAD21, RAD21L, or REC8), and a stromal antigen protein (STAG1, 2, or 3). STAG3 is exclusively expressed during meiosis, and is the predominant STAG protein component of cohesin complexes in primary spermatocytes from mouse, interacting directly with each α-kleisin subunit. REC8 and RAD21L are also meiosis-specific cohesin components. Stag3 mutant spermatocytes arrest in early prophase ("zygotene-like" stage), displaying failed homolog synapsis and persistent DNA damage, as a result of unstable loading of cohesin onto the chromosome axes. Interestingly, Rec8, Rad21L double mutants resulted in an earlier "leptotene-like" arrest, accompanied by complete absence of STAG3 loading. To assess genetic interactions between STAG3 and α-kleisin subunits RAD21L and REC8, our lab generated Stag3, Rad21L, and Stag3, Rec8 double knockout mice, and compared them to the Rec8, Rad21L double mutant. These double mutants are phenotypically distinct from one another, and more severe than each single knockout mutant with regards to chromosome axis formation, cohesin loading, and sister chromatid cohesion. The Stag3, Rad21L, and Stag3, Rec8 double mutants both progress further into prophase I than the Rec8, Rad21L double mutant. Our genetic analysis demonstrates that cohesins containing STAG3 and REC8 are the main complex required for centromeric cohesion, and RAD21L cohesins are required for normal clustering of pericentromeric heterochromatin. Furthermore, the STAG3/REC8 and STAG3/RAD21L cohesins are the primary cohesins required for axis formation.
Binding Sites in the EFG1 Promoter for Transcription Factors in a Proposed Regulatory Network: A Functional Analysis in the White and Opaque Phases of Candida albicans
Pujol, C., Srikantha, T., Park, Y.-N., Daniels, K. J., Soll, D. R.
In Candida albicans the transcription factor Efg1, which is differentially expressed in the white phase of the white-opaque transition, is essential for expression of the white phenotype. It is one of six transcription factors included in a proposed interactive transcription network regulating white-opaque switching and maintenance of the alternative phenotypes. Ten sites were identified in the EFG1 promoter that differentially bind one or more of the network transcription factors in the white and/or opaque phase. To explore the functionality of these binding sites in the differential expression of EFG1, we generated targeted deletions of each of the 10 binding sites, combinatorial deletions, and regional deletions using a Renilla reniformis luciferase reporter system. Individually targeted deletion of only four of the 10 sites had minor effects consistent with differential expression of EFG1, and only in the opaque phase. Alternative explanations are considered.
Chromosome-Wide Impacts on the Expression of Incompatibilities in Hybrids of Tigriopus californicus
Willett, C. S., Lima, T. G., Kovaleva, I., Hatfield, L.
Chromosome rearrangements such as inversions have been recognized previously as contributing to reproductive isolation by maintaining alleles together that jointly contribute to deleterious genetic interactions and postzygotic reproductive isolation. In this study, an impact of potential incompatibilities merely residing on the same chromosome was found in crosses of populations of the copepod Tigriopus californicus. When genetically divergent populations of this copepod are crossed, hybrids show reduced fitness, and deviations from expected genotypic ratios can be used to determine regions of the genome involved in deleterious interactions. In this study, a set of markers was genotyped for a cross of two populations of T. californicus, and these markers show widespread deviations from Mendelian expectations, with entire chromosomes showing marked skew. Despite the importance of mtDNA/nuclear interactions in incompatibilities in this system in previous studies, in these crosses the expected patterns stemming from these interactions are not widely apparent. Females lack recombination in this species, and a striking difference is observed between male and female backcrosses. This suggests that the maintenance of multiple loci on individual chromosomes can enable some incompatibilities, perhaps playing a similar role in the initial rounds of hybridization to chromosomal rearrangements in preserving sets of alleles together that contribute to incompatibilities. Finally, it was observed that candidate pairs of incompatibility regions are not consistently interacting across replicates or subsets of these crosses, despite the repeatability of the deviations at many of the single loci themselves, suggesting that more complicated models of Dobzhansky-Muller incompatibilities may need to be considered.
N-Ethyl-N-Nitrosourea (ENU) Mutagenesis Reveals an Intronic Residue Critical for Caenorhabditis elegans 3' Splice Site Function in Vivo
Itani, O. A., Flibotte, S., Dumas, K. J., Guo, C., Blumenthal, T., Hu, P. J.
Metazoan introns contain a polypyrimidine tract immediately upstream of the AG dinucleotide that defines the 3' splice site. In the nematode Caenorhabditis elegans, 3' splice sites are characterized by a highly conserved UUUUCAG/R octamer motif. While the conservation of pyrimidines in this motif is strongly suggestive of their importance in pre-mRNA splicing, in vivo evidence in support of this is lacking. In an N-ethyl-N-nitrosourea (ENU) mutagenesis screen in Caenorhabditis elegans, we have isolated a strain containing a point mutation in the octamer motif of a 3' splice site in the daf-12 gene. This mutation, a single base T-to-G transversion at the -5 position relative to the splice site, causes a strong daf-12 loss-of-function phenotype by abrogating splicing. The resulting transcript is predicted to encode a truncated DAF-12 protein generated by translation into the retained intron, which contains an in-frame stop codon. Other than the perfectly conserved AG dinucleotide at the site of splicing, G at the –5 position of the octamer motif is the most uncommon base in C. elegans 3' splice sites, occurring at closely paired sites where the better match to the splicing consensus is a few bases downstream. Our results highlight both the biological importance of the highly conserved –5 uridine residue in the C. elegans 3' splice site octamer motif as well as the utility of using ENU as a mutagen to study the function of polypyrimidine tracts and other AU- or AT-rich motifs in vivo.
Genome Sequence and Analysis of a Stress-Tolerant, Wild-Derived Strain of Saccharomyces cerevisiae Used in Biofuels Research
McIlwain, S. J., Peris, D., Sardi, M., Moskvin, O. V., Zhan, F., Myers, K. S., Riley, N. M., Buzzell, A., Parreiras, L. S., Ong, I. M., Landick, R., Coon, J. J., Gasch, A. P., Sato, T. K., Hittinger, C. T.
The genome sequences of more than 100 strains of the yeast Saccharomyces cerevisiae have been published. Unfortunately, most of these genome assemblies contain dozens to hundreds of gaps at repetitive sequences, including transposable elements, tRNAs, and subtelomeric regions, which is where novel genes generally reside. Relatively few strains have been chosen for genome sequencing based on their biofuel production potential, leaving an additional knowledge gap. Here, we describe the nearly complete genome sequence of GLBRCY22-3 (Y22-3), a strain of S. cerevisiae derived from the stress-tolerant wild strain NRRL YB-210 and subsequently engineered for xylose metabolism. After benchmarking several genome assembly approaches, we developed a pipeline to integrate Pacific Biosciences (PacBio) and Illumina sequencing data and achieved one of the highest quality genome assemblies for any S. cerevisiae strain. Specifically, the contig N50 is 693 kbp, and the sequences of most chromosomes, the mitochondrial genome, and the 2-micron plasmid are complete. Our annotation predicts 92 genes that are not present in the reference genome of the laboratory strain S288c, over 70% of which were expressed. We predicted functions for 43 of these genes, 28 of which were previously uncharacterized and unnamed. Remarkably, many of these genes are predicted to be involved in stress tolerance and carbon metabolism and are shared with a Brazilian bioethanol production strain, even though the strains differ dramatically at most genetic loci. The Y22-3 genome sequence provides an exceptionally high-quality resource for basic and applied research in bioenergy and genetics.
Fine-Scale Crossover Rate Variation on the Caenorhabditis elegans X Chromosome
Bernstein, M. R., Rockman, M. V.
Meiotic recombination creates genotypic diversity within species. Recombination rates vary substantially across taxa, and the distribution of crossovers can differ significantly among populations and between sexes. Crossover locations within species have been found to vary by chromosome and by position within chromosomes, where most crossover events occur in small regions known as recombination hotspots. However, several species appear to lack hotspots despite significant crossover heterogeneity. The nematode Caenorhabditis elegans was previously found to have the least fine-scale variation in crossover distribution among organisms studied to date. It is unclear whether this pattern extends to the X chromosome given its unique compaction through the pachytene stage of meiotic prophase in hermaphrodites. We generated 798 recombinant nested near-isogenic lines (NILs) with crossovers in a 1.41 Mb region on the left arm of the X chromosome to determine if its recombination landscape is similar to that of the autosomes. We find that the fine-scale variation in crossover rate is lower than that of other model species, and is inconsistent with hotspots. The relationship of genomic features to crossover rate is dependent on scale, with GC content, histone modifications, and nucleosome occupancy being negatively associated with crossovers. We also find that the abundances of 4- to 6-bp DNA motifs significantly explain crossover density. These results are consistent with recombination occurring at unevenly distributed sites of open chromatin.
A Comprehensive Toolbox for Genome Editing in Cultured Drosophila melanogaster Cells
Kunzelmann, S., Bottcher, R., Schmidts, I., Forstemann, K.
Custom genome editing has become an essential element of molecular biology. In particular, the generation of fusion constructs with epitope tags or fluorescent proteins at the genomic locus facilitates the analysis of protein expression, localization, and interaction partners at physiologic levels. Following up on our initial publication, we now describe a considerably simplified, more efficient, and readily scalable experimental workflow for PCR-based genome editing in cultured Drosophila melanogaster cells. Our analysis at the act5C locus suggests that PCR-based homology arms of 60 bp are sufficient to reach targeting efficiencies of up to 80% after selection; extension to 80 bp (PCR) or 500 bp (targeting vector) did not further improve the yield. We have expanded our targeting system to N-terminal epitope tags; this also allows the generation of cell populations with heterologous expression control of the tagged locus via the copper-inducible mtnDE promoter. We present detailed, quantitative data on editing efficiencies for several genomic loci that may serve as positive controls or benchmarks in other laboratories. While our first PCR-based editing approach offered only blasticidin-resistance for selection, we now introduce puromycin-resistance as a second, independent selection marker; it is thus possible to edit two loci (e.g., for coimmunoprecipitation) without marker removal. Finally, we describe a modified FLP recombinase expression plasmid that improves the efficiency of marker cassette FLP-out. In summary, our technique and reagents enable a flexible, robust, and cloning-free genome editing approach that can be parallelized for scale-up.
Site-Directed Genome Knockout in Chicken Cell Line and Embryos Can Use CRISPR/Cas Gene Editing Technology
Zuo, Q., Wang, Y., Cheng, S., Lian, C., Tang, B., Wang, F., Lu, Z., Ji, Y., Zhao, R., Zhang, W., Jin, K., Song, J., Zhang, Y., Li, B.
The present study established an efficient genome editing approach for the construction of stable transgenic cell lines of the domestic chicken (Gallus gallus domesticus). Our objectives were to facilitate the breeding of high-yield, high-quality chicken strains, and to investigate gene function in chicken stem cells. Three guide RNA (gRNAs) were designed to knockout the C2EIP gene, and knockout efficiency was evaluated in DF-1 chicken fibroblasts and chicken ESCs using the luciferase single-strand annealing (SSA) recombination assay, T7 endonuclease I (T7EI) assay, and TA clone sequencing. In addition, the polyethylenimine-encapsulated Cas9/gRNA plasmid was injected into fresh fertilized eggs. At 4.5 d later, frozen sections of the embryos were prepared, and knockout efficiency was evaluated by the T7EI assay. SSA assay results showed that luciferase activity of the vector expressing gRNA-3 was double that of the control. Results of the T7EI assay and TA clone sequencing indicated that Cas9/gRNA vector-mediated gene knockdown efficiency was approximately 27% in both DF-1 cells and ESCs. The CRISPR/Cas9 vector was also expressed in chicken embryos, resulting in gene knockdown in three of the 20 embryos (gene knockdown efficiency 15%). Taken together, our results indicate that the CRISPR/Cas9 system can mediate stable gene knockdown at the cell and embryo levels in domestic chickens.
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Patrick Cockburn: How Neoliberalism and Cold War Obsessions Fractured the Middle East - How neoliberalism, the resource curse, and military interventions by Washington and its allies, profoundly destabilized the Middle East.
37 minutes ago
Addressing threats to health care's core values, especially those stemming from concentration and abuse of power. Advocating for accountability, integrity, transparency, honesty and ethics in leadership and governance of health care.
Who are considered the best skeptical bloggers about Ethics Code?


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Posted on: November 18th in Destinations, Europe, Travel, Travel Photography by Graham Padmore.
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Photo: Joshua Strang, USAF
The northern lights or aurora borealis have fascinated many for thousands of years. A naturally occurring phenomenon, the lights appear as solar wind particles collide with air molecules in the Earth's atmosphere, and emit excess energy as light. The winter months provide the best conditions to see them in the northern regions of Iceland, Canada, Finland, Sweden and Norway. You're not always guaranteed to see them, but on a clear evening, far away from natural lights, and as far north as possible, you could witness this incredible display of nature. These are the beautiful Northern Lights in Pictures:
Where to stay
- Sweden: Icehotel – Built entirely from snow and ice, a stylish hotel north of the Arctic Circle in Lapland
- Iceland: Hotel Rangá – A member of the Special Hotels Of The World, a luxury resort 1hr from Reykjavik
- Finland: Holiday Club Hotel Saariselka – The northernmost spa hotel in Europe
- Canada: Explorer Hotel – In northwest Canada, 500km from the Arctic Circle
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Here's a video briefing the East European Imperial Alliance, the main army.
Next, a video on Gallia which introduces several minor characters.
A look at the Nameless' mobile camp and customizing your units and tanks:
Lastly, this is an overview of the Nameless (the main squad) and its characters.
Valkyria Chronicles 3 will be out in Japan on January 27th.
Wild Arms 2 July 3 • 9:00am PDT/12:00pm EDT
Lunar 2: Eternal Blue Continues! Every Monday • 5:00pm PDT/8:00pm EDT
Zero Time Dilemma Every Night! • 5:00pm PDT/8:00pm EDT
Grandia II Continues! Every Wednesday • 2:00pm PDT/5:00pm EDT


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NEW YORK (AP) — In its first two years of operation, the small group of lawyers handling a compensation program for people with health problems related to the Sept. 11 attacks has been bogged down in bureaucracy and has issued decisions to only 112 people, according to a report released Friday by the fund's chief administrator.
In the report, Sept. 11th Victim Compensation Fund special master Sheila Birnbaum revealed that only a trickle of money has gone out. Of the $2.78 billion appropriated by Congress, only $27.2 million has been awarded. Little of that cash has actually reached applicants.
Now the fund may face an even more daunting challenge: processing a deluge of 55,000 applications, many of which came in over the summer and early fall as people exposed to dust at the World Trade Center raced to beat an October deadline.
The slow pace has upset some advocates for the sick. It also raises questions about whether the fund has the resources it needs to handle the mountain of applications from police officers, firefighters, construction and cleanup workers, and other people who were caught in the dust cloud when the twin towers collapsed, or who worked on the burning debris pile.
Birnbaum recently boosted staff to relieve the bottleneck. The fund now employs 75 people, up from 31 a year ago.
In the report, she said the fund is working to streamline the review process. In addition to hiring more people, it has been refining the computerized system used to review and track claims. It has brought in physicians to help review medical claims.
Birnbaum said one big hurdle has been difficulty gathering required documents.
Applicants are required to submit paperwork showing that they were present at the World Trade Center site, or responded to the Pentagon or the field in western Pennsylvania where terrorists crashed a hijacked jet.
They also have to provide certain medical records, or alternatively enroll in a special treatment program for people sickened by the attacks, and get a doctor there to verify that their illness could be related to the attacks.
Finally, applicants also have to document any economic losses they suffered as a result of their illness, since that will form the basis of their compensation payment.
Of the many applicants, only around 2,500 have submitted complete eligibility forms, and only a few hundred have sent in all the documents required to issue a compensation decision the report said.