Pure translation display

Pure translation display
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DOI:
10.1016/j.ab.2004.06.028
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发表时间:
2004-10-15
影响因子:
2.9
通讯作者:
Blacklow, SC
Blacklow, SC
中科院分区:
生物学4区
文献类型:
--
作者:
Forster, AC;Cornish, VW;Blacklow, SC

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Methods such as monoclonal antibody technology, phage display, and ribosome display provide genetic routes to the selection of proteins and peptides with desired properties. However, extension to polymers of unnatural amino acids is problematic because the translation step is always performed in vivo or in crude extracts in the face of competition from natural amino acids. Here, we address this restriction using a pure translation system in which aminoacyl-tRNA synthetases and other competitors are deliberately omitted. First, we show that such a simplified system can synthesize long potypeptides. Second, we demonstrate "pure translation display" by selecting from an mRNA library only those mRNAs that encode a selectable unnatural amino acid upstream of a peptide spacer sequence long enough to span the ribosome tunnel. Pure translation display should enable the directed evolution of peptide analogs with desirable catalytic or pharmacological properties. (C) 2004 Elsevier Inc. All rights reserved.