anuvaada shaastram

translatomics anedi oka kanam ledhaa jeevilo churukuga anuvadinchabadutunna anni open reeding framela (orfs) adhyayanam. eee rfs sekarananu translatome antaaru. oka kanam yokka translatomnu vargeekarinchadam anedi kanam loe churukuga unde jeeva maargaala shrenipai antardrushtini istundi. molecular biology yokka kendra siddhaantam prakaram, oka kanamlooni dna rnanu utpatti cheyadaaniki lipyantareekarinchabadutundi, idhi taruvaata proteinnu utpatti cheyadaaniki anuvadinchabadutundi. oka jeevi yokka janyuvulo vaelaadi proteins encode cheyabadataayi, mariyu oka kanamlo unde proteins kanam yokka jeevitaaniki maddatugaa aneka vidhulanu sahakaaramtho nirvahistaayi. ottidi ledhaa abhivruddhilo nirdishta kaala binduvulu vanti vividha paristhitulalo, kanam churukuga undataaniki vividha jeeva margalu avasaram kaavacchu, anduvalla proteinla yokka vibhina sekarana avasaram. antargata mariyu paryaavarana paristhitulanu batti, okesari tayaaru cheyabadutunna proteinla sekarana maarutuu untundi. kriyaaseela anuvaadam chese rfs yokka eee sekarana yokka "snapshot" teesukoovadaaniki anuvaada saanketikatalanu upayoginchavacchu, idhi prastuta paristhitulalo kanam yea jeeva maargaalanu sakriyam chestondi anae samacharanni istundi.
saadhaaranamgaa, ribozome profiling technic translatome samacharanni pondadaaniki upayoginchabadutundi. single-sell ribozome profilingtho sahaa iteevali purogatulu, eee adhyayanala spashtatanu gananeeyamgaa meruguparichayi, parisoedhakulu vyaktigata kanaala sthaayilo anuvaadampai antardrushtulanu pondadaaniki veelu kalpinchaayi. idhi bhinnamaina kana janaabhaaku chaalaa mukhyam, ikkada mottam sthuula kolatalu protein samshlaeshanhaloo mukhyamaina kana-nundi-kana vyatyaasaalanu kappipuchavacchu. itara paddhatulu polysome profiling, puurti-nidivi anuvaadam mrna profiling (arnc-sec), mariyu anuvaadam ribozome affinity purification (trop-sec). transcriptome kakunda, konni janyuvula vyakteekarana sthaayini anchanaa vaeyadaaniki translatome marinta khachitamaina ujjaayimpu, endukante proteom mariyu translatome madhya sahasambandham transcriptome mariyu proteom madhya sahasambandham kante ekkuvagaa untundi.
charitra.
maanava janyuparamaina project poortayye samayaniki janyusaastram yokka rangam janyuvula vidhulanu nirnayinche disagaa thana drushtini maarchukundi. indulo kanaalalo rna mariyu proteins vanti jeeva padaarthaala itara sekaranalanu jabita cheyadam untundi. eee padaarthaala sekaranalanu-oms anie pilicheevaaru, idhi maanava janyuvu yokka kramam chuttoo vistrutamaina utsahanni rekettinchindi. translatome anae padaanni modatisaarigaa modatisaarigaa greenbaum mariyu itarulu pratipaadinchaaru. translatome anedi proteomloni proteinla saapeksha parimaanaalanu vivarinchadaaniki uddesinchabadindi. ippudu translatome anae padam saadhaaranamgaa kanamlo churukuga srushtinchabadutunna proteinla sekarananu suuchistumdi. anuvaadasaastram, adhokaranam kalayikatho, vividha paristhitulalo proteomku nikara maarpunu vivarinchadam lakshyamgaa pettukundi.
omiksku auchityam mariyu sahakaram.
janyuparamaina adhyayanam ledhaa oka jeevilo janyu padaarthaala sekarana anedi janyusaastram yokka lakshyam. janyusaastra upakshetraalu ledhaa transcriptomics mariyu proteomics vanti itara-omics, vividha paristhitulalo janyuvu yokka utpattulanu (rna mariyu proteins vantivi) lekkinchadam dwara janyuvu paniteerunu vargeekarinchadam lakshyamgaa pettukunnayi. alaa cheyadamlo, omics janyu vyakteekarana yokka vividha sthaayila niyantrana mariyu anduvalla janyu paniteeru gurinchi antardrushtini pondutaaru. ayithe, eee kshetraalu ippatike erpadina jeeva anuvulanu vargeekaristaayi. konni sandarbhaallo, rna ledhaa protein samruddhi paniteerunu pratibimbimchadu endukante eee jeeva anuvulu vaegamgaa ksheeninchavacchu ledhaa avi praarambhamlo samslaeshanha cheyabadina chaalaa kaalam tarvaata kanamlo undavacchu. proteomnu adhyayanam cheyadaaniki proteomics paddhatulanu upayogistunnappudu, anuvaadaanantara maarpu mariyu protein ksheenata sthaayilo protein samruddhini niyantrinchadam munupati niyantrana prakriyalanu aspashtamgaa cheyavachu. endukante cellular functionlu tarachugaa anuvaada sthaayilo niyantrinchabadataayi, antey transcriptome ellappuduu janyuvu paniteerunu pratibimbimchadu, translatomnu adhyayanam cheyadaaniki translatomics paddhatulanu upayoginchadam anedi transcriptomics ledhaa proteomics adhyayanaalalo aspashtamgaa unde janyuvu paniteeru yokka niyantrananu gamaninchadaaniki veelu kalpistundi.
paddhatulu.
mrna messenger rnani anuvadistondi.
chaalaa anuvaada shaastra paddhatulu raibojomlatho sanklishtamgaa unde mrunaa lanu vargeekarinchadampai drushti pedataayi, anduvalla anuvaadam cheyabadataayi.
polysome profiling.
polysome profiling anedi okati ledhaa anthakante ekkuva mrnla anuvaada sthaayini vargeekarinchadaaniki upayoginche oka saanketikata. atyanta anuvadinchabadina mrna oka palisomga undhi, antey idhi bahula raibojomlatho sanklishtamgaa untundi. diguva sthaayilalo anuvadinchabadina mrnlu takkuva raibojomlatho sanklishtamgaa untaayi. polysome profilinglo, parimaanam aadhaaramgaa sell laisetloni molecular complexelanu vaeru cheyadaaniki sukroj pravanata upayoginchabadutundi. niluvu varusa nundi bhinnalu kramam ledhaa itara paddhatula dwara vislaeshimchabadataayi. takkuva mariyu ekkuva paramaanu baruvu gala bhinnalalo daani gurtimpu mariyu samruddhi aadhaaramgaa mrnla anuvaada raetu nirnayinchabadutundi.
rnc-sec.
puurti podavu anuvaadam chese mrna (arnc-sec) loe sukroj kushanpai liceted namuunaanu centrifugation chestaaru. idhi ribozome-nacent chain complexes (arnc) nuu free mrnlu mariyu itara kana bhagala nundi vaeru cheyadaaniki anumatistundi. arnclu centrifugationlo oka gulikanu yerparustaayi, idhi tadupari vislaeshanha kosam sekarinchabadutundi. eee arnclalo anuvadinchabadutunna mrnanu kramabaddheekarinchavachhu, aaa samayamlo anuvadinchabadutunna mrnlanu gurtinchadaaniki mariyu parimaanam cheyadaaniki veelu kalpistundi. ayithe, arnc-mrna complexelu pelusugaa untaayi, ivi ribozomelu mrnla nundi vidipovadaaniki mariyu mrnla ksheenataku daariteestaayi, sekarinchina phalitaalanu pakshapaatam chestaayi.
raibo-sec/ribozome profiling/ribozome footprinting.
ribozome profilinglo, paalisomlatoe sahaa cellular mrna ribonucleuslaku lobadi untundi, ivi rnanu cheelche enjaimlu. raibojomlatho bandhinchabadina rna anuvulalo aaa sthaanaalu jeernakriya nundi rakshinchabadataayi. ribonucles karyakalapalanu nilipivesina taruvaata, eee rakshita pradaesaalanu tirigi pondavacchu mariyu kramabaddheekarinchavachhu. anduvalla raibo-sec nundi pondina kramaalu churukuga anuvadinchabadutunna mrunaa sakalaalanu suuchistaayi.
trop-sec.
oka kanajaalam lopala ledhaa itara rakaala kanaalalo oka nirdishta kana rakamlo churukuga anuvadinchabadutunna mrunaa lanu gurtinchadaaniki trop-sec upayoginchabadutundi. aakupaccha fluorescent protein vanti epitope tagku anusandhaaninchabadina ribosomal sub unitnu vyakteekarinchadaaniki sell rakam aasakti roopondinchabadindi. sell lysis taruvaata, epitopnu lakshyamgaa chesukune pratiroedhakaalu fusion proteinlanu kaligi unna ribozomelaku kattubadi unna mrunaa lanu vaeru cheyadaaniki upayoginchabadataayi. eee rna appudu cdnaga maarchabadutundi mariyu kramabaddheekarinchabadutundi. eee saanketikata pratyaekamgaa aasaktigala kana rakamlo anuvadinchabadutunna mrnlanu gurtistundi.
navjata polypeptides.
madata rashtraalu.
maas spectrometry.
maas spectrometry paddhatulu navjata polypeptides madatanu nirnayinchalevu. prastuta paddhatulu edhee kanamlo prapanchavyaaptamgaa navjata polypeptides yokka madata sthithulanu pariseelinchavu. vyaktigata navjata polypeptides yokka madata sthithini pariseelinchadaaniki paddhatulu unnaayi.
oka paddhati takkuva ushnograta vadda navjata peptidenu cheelchadaaniki anirdishta proteasenu upayogistundi. protease vippabadina, anuvaina praantaalanu cheelchagaladu, conny gattiga madathapette praantaalanu kattirinchadu. cheelika yokka utpattulanu appudu vaeru chesi, navjata peptide yokka madata praantaalanu nirnayinchadaaniki adhyayanam cheyavachu.
nuclear magnetic resonance.
navjata polypeptide yokka puurti nirmaanaanni vislaeshimchadaaniki, nuclear magnetic resonance (nmr) upayoginchabadutundi. nmr draavanamlooni anuvula dynamic veekshanhanu anumatistundi mariyu anduvalla ribozome-nacent chain complexelalo (arnc) upayoginchavacchu. ribozomela labeling anedi nmr daetaanu anumaanita ribozome signal kosam filter cheyadaaniki mariyu navjata polypeptide yokka signalnu gurtinchadaaniki anumatistundi.
gurtimpu mariyu parimaanam.
navjata polypeptides nirmaanam prastutam prapancha sthaayilo gurtinchalenidi ayinappatikee, gurtimpu mariyu parimaanam ledhu.
kana samskrutilo amaino aamlaala dwara sthiramaina isotope labeling.
oka paddhati kana samskruti (silac) loe amaino aamlaala dwara sthiramaina isotope labeling yokka vaividhyaanni upayogistundi. silak parimaanaanni anumatinchadaaniki sthiramaina isotopulatho proteinlanu label chestundi, parimaanam kosam label cheyabadina mariyu label cheyani peptidelanu polchi chustundi. pulls silac (silac), pulls samayamlo srushtinchabadina peptidelanu matrame label cheyadaaniki anumatistundi. siddhaantaparamgaa, idhi parimaanam kosam navjata peptidelanu matrame sangrahinchadaaniki anumatistundi. ayithe, silakku khachitamaina parimaanam kosam label cheyabadina mariyu label cheyani proteinla yokka saaruupya sthaayilu avasaram. anduvalla, sailak pulls anuvaada prakriya kante chaalaa ekkuva kaalam panicheyaali, tadwara navjata peptidela parimaanam sarikaadu.
bio-orthogonal/quantitative naan-canonical amaino assid tagging.
bio-orthogonal/quantitative naan-canonical amaino assid tagging (boncat/quancat) proteinlanu tagg cheyadaaniki azidohomalanine (aahaa) nuu upayogistundi. idhi ms kosam kottagaa srushtinchina proteinlanu vaeruchaeyadaaniki anumatistundi. ayithe, aahanu upayoginchadaaniki kanaantara methionin yokka mundastu tolagimpu mariyu aahanu pravesapettadam avasaram, idhi kanaanni nokki chebutundi mariyu lopala anuvaada gatiseelatanu maarchagaladu. sailak maadirigaane, aahaa paddhatulaku podavaina pulls avasaram, tadwara navjata peptidesnu lekkinchadamlo daani saamarthyaanni parimitam chestundi.
puromycin-sambandhita navjata golusu proteomics.
puromycin-sambandhita nacent chain protemics (punch-pii) ms kosam nacent polypeptidesnu sangrahinchadaaniki puromycin-biotine lebulnu upayogistundi. idhi kana prakriyaku bhangam kaliginchanappatikii, navjata peptidelanu gurtinchadamlo itara paddhatula kante takkuva sunnitamainadi. eee parimaanaatmaka paddhathulatho paatu, itara ms aadhaarita paddhatulapai pani jarugutondi.
mrna ksheenata.
anuvaada prakriyanu niyantrinchadamlo mrna ksheenata kooda mukhyamaina paatra poeshistumdi.
kshayam yokka vidhaanaalanu anveshinchadaaniki, uncape cheyani mariyu cheelika transcripts (gmuct) yokka janyu-wide mapping, rna chivarala samaantara vislaeshanha (paerae), mariyu degradum sequencing, ksheeninchina mrnlanu kramam cheyadaaniki prakaasam sequencing platfam yokka upayogistaayi ligesnu upayogistaayi. upayogistaayi ligase oka uchita 5 'monophosphate thoo rna meeda ligats chestundi. paripakva mrunaasayam 5 'podupu kaligi unnanduna, avi uparitalaalugaa bandhinchabadavu, tadwara sirachchaedam cheyabadi, adhokaranam chendutunna mrunaashalam bandhinchabadataayi.
5′-monophosphorylated ends sequencing (5pseq) paripakva mrna mariyu adhokaranam chendina utpattulu rendinti sequencingnu anumatinchadaaniki kappabadina mariyu sirachchaedam cheyabadina sequencelu rendintinee sangrahistundi. idhi mrna ksheenata utpattulanu gurtinchadamlo sahaayapadutundi mariyu ribozome stallingnu adhyayanam cheyadamlo upayogapadutundi.
eee paddhatulu 5 'nundi 3' ksheenata, mirna-madhyavartitva cheelika mariyu ardhamleni madhyavartitva mrunaa kshayam gurinchi adhyayanam chestaayi, conny 3 'nundi 5' ksheenata mariyu itara ksheenata vidhaanaalanu kolavalevu.
vivo anuvaada tracking loe.
paina perkonna paddhatulaku kanaala lysis avasaram, anduvalla jeeva kanaalalo nirvahinchabadadu. single molecule fluorescence resonance energy transfer (smfret) mariyu nacent chain tracking (nct) translational activityni track cheyadaaniki fluorescensenu upayogistaayi. rendu paddhatulu oke mrnlpy polypeptides yokka podavunu track chestaayi. ayithe, yea saanketikata kooda adhika nirgamaamsaanni andinchalevu.
tranome.
anuvaadamlo badilee rna (trna) oka mukhyamaina bhaagam. traina luu mrna nii chadivi, ribozome sameekarinche amaino aamlaalanu polypeptide loki teesukuvastaayi. anduvalla, trinaala samruddhi mariyu rakalu protein samslaeshanha vegampai pedda prabhavanni chuuputaayi. konni trana jaatulu oke nucleotide dwara matrame bhinnamgaa untaayi, tranalu itara tranalatho chaalaa pooli untaayi. idhi, saaruupya dviteeya mariyu trutheeya nirmaanaalatoo kalisi vividha trana jaatulanu vaeru cheyadam kashtataram chestundi.
zell electrophoresis.
jaatulanu electrophoresis anedi tranalanu vaeru cheyadaaniki upayoginche oka praamaanika paddhati. praarambhamlo, tranalu 7 mea eurialo vikruteekarinchabadataayi mariyu modati zell parimaanamlo vaeru cheyabadataayi. 4 mea euria rendava zell parimaanamlo adanapu vibhajana kosam pakshika refoldingnu anumatistundi. eee paddhati e loe 48 setlugaa vaeru cheyadaaniki anumatinchindi. kooli mariyu 30 inn bii. subtilis conny parimita spashtata kaligi untundi. 269 elukala tranalaku 62 macchalu matrame kanipinchadamtho, pedda sankhyalo vividha trana jaatulanu jaatulanu electrophoresis dwara puurtigaa vaeru cheyalemu.
liquid chromatography.
adhika paniteeru gala drava chromatographyni aminoacylated trna isooxeptorla aadhaaramgaa tranalanu vaeru cheyadaaniki upayoginchavacchu. eee paddhati trna jaatulanu puurtigaa vaeru cheyadu mariyu kodanla madhya thedaanu gurtinchadu, ayinappatikee idhi ippatikee vividha kana rekhala madhya parimaanaatmaka vyatyaasaalanu kanugonagaladu.
maas spectrometry.
pratyaekamaina endonucles jeerna utpattula aadhaaramgaa tranalanu vaeru cheyadaaniki maas spectrometry (ms) nuu upayoginchavacchu. ayithe, idhi 30 trana jaatula misramaalato parimita spashtatanu kaligi undhi mariyu trana yokka pedda samoohaalalo ms kante mundu bhinnam avasaram. idhi denovo trna jaatulanu gurtinchadaaniki kooda upayoginchabadadu endukante dheeniki trna jaatula jeerna vidhaanaala gurinchi mundastu ghnaanam avasaram.
micro arres.
hybridisation aadhaarita microarrelu fluorescent probnu jodinchadaaniki tranalalo 3 'pakka samrakshinchabadina kramaanni upayogistaayi. trana podavunu kappi unche nucleotide podavaina probes, tranalanu bandhinchadaaniki upayoginchabadataayi. kaneesam 8 bays differencelu unna tranalanu microarelatho vaeru cheyagalaru, conny chinna differencelu unna tranalu oke probto bandhistaayi.
parimaanaatmaka riverse transcription.
quantitative riverse transcription, real time pcr (qrt-pcr) anedi tranomnu gurtinchadaaniki mariyu lekkinchadaaniki maroka maargam. anni trna jaatulaku riverse transcription priming kosam samrakshinchabadina 3 'pakka sequence kosam primer upayoginchabadutundi. trna nucleotidela yokka bhaarii maarpu mariyu trana anuvu yokka sthirathvam oka samasya kaavacchu, kabatti veetini edurkovadaniki adhika ushnogratalu mariyu demethylation upayoginchabaddaayi. sequencinglo bhaarii methylation prerepanalanu nirodhinchadaaniki kooda demethylation upayoginchabadindi.
raibo-sec vantidi.
anuvaadaaniki marinta daggaragaa trana paatranu gamaninchadaaniki raibo-trana-sec abhivruddhi cheyabadindi. raibo-sec maadirigaane, raibo-trna-sec, kramabaddheekaranaku mundu library tayari kosam ribozomelalo trna anuvulanu sangrahistundi.

