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Biotech / Medical : Oxford GlycoSciences Plc -- Ignore unavailable to you. Want to Upgrade?

To: John McCarthy who wrote (344)	2/20/2003 3:33:51 AM
From: nigel bates	Read Replies (1) \| Respond to of 469

Thanks. Could be one of the motivations for the merger? ...Such a "ribosome display" approach has a number of disadvantages, including the fact that the complexes obtained also include all elements of the protein synthesis machinery, i.e. ribosomes with all their associated RNAs and proteins. This not only depletes the translation reaction but also results in a very high background and large number of unrelated proteins linked to the mRNA. Xu et al [6] have produced intermediate {mRNA-DNA-adapter-ribosome-Protein} complexes where a puromycin-labelled DNA adapter, separately ligated to RNA molecules, covalently links to a nascent protein chain in a sequence-independent manner (an "mRNA display" approach, [6]). Such a modification results in covalent {mRNA-protein} complexes, which lack bulky ribosomes, but involve a high degree of non-specific crosslinking of the RNA to ribosomal proteins. Ligation of a puromycin-modified DNA to mRNA requires an additional step, which makes the whole procedure significantly longer especially if a few rounds of subsequent amplification and selection are required. A variation of RNA-protein complex production using puromycin was also reported by Roberts and Szostak, and by Liu et al [7,8] respectively. All the methods reported so far result in the production of covalently crosslinked protein-RNA hybrids and/or complexes containing bulky ribosomes or requiring multi-step processes and excessive RNA handling in order to make protein-DNA complexes...