One of the most challenging requirements associated with combinatorial chemistry is the recovery of sequence information of the oligonucleotide or peptide selected by the screening assay. A method1 has been developed to generate a fusion product between mRNA and the polypeptide it encodes using in vitro translation of synthetic RNAs 3'-labeled with puromycin, an antibiotic that mimics transfer RNA. Puromycin binds in the ribosome's A site, forms a peptide bond with the growing peptide chain, and blocks further peptide elongation. By linking puromycin to mRNA, a peptide-RNA fusion product results from the translation of the message linking the encoding mRNA with its peptide product.
Coupling: This support should be used in a manner identical to normal protected nucleoside support since it contains the DMT group.
Deprotection: Deprotect using ammonium hydroxide as required by the nucleobases. Note, using AMA will lead to degradation of the puromycin and is not recommended.
Controlled room temperature or lower, dry
1 R.W. Roberts and J.W. Szostak, Proc. Natl. Acad. Sci. USA, 1997, 94, 12297-302.