be avoided, particularly in washing glassware used for lyophilization. A feature of the native chemical ligation technique is that the product polypeptide chain contains cysteine at the site of ligation. The cysteine at the ligation site can be desulfurized to alanine, thus extending the range of possible ligation sites to include alanine residues. Other beta-thiol containing amino acids can used for native chemical ligation, followed by desulfurization. Alternatively, thiol-containing ligation auxiliaries can be used that mimic an N-terminal cysteine for the ligation reaction, but which can be removed after synthesis. The use of thiol-containing auxiliaries may not be as effective as ligation at a Cys residue. Native chemical ligation can also be performed with an N-terminal selenocysteine residue. Polypeptide C-terminal thioesters produced by recombinant DNA techniques can be reacted with an N-terminal Cys containing polypeptide by the same native ligation chemistry to provide very large semi-synthetic proteins. Native chemical ligation of this kind using a recombinant polypeptide segment is known as Expressed Protein Ligation. Similarly, a recombinant protein containing an N-terminal Cys can be reacted with a synthetic polypeptide thioester. Thus, native chemical ligation can be used to introduce chemically synthesized segments into recombinant proteins, regardless of size. See also Intein KAHA Ligation Peptide synthesis Protein synthesis SEA Native Peptide Ligation References Further reading Peptides