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In vitro virus: Bonding of mRNA bearing puromycin at the 3′‐terminal end to the C‐terminal end of its encoded protein on the ribosome in vitro
Author(s) -
Naoto Nemoto,
Etsuko MiyamotoSato,
Yuzuru Husimi,
Hiroshi Yanagawa
Publication year - 1997
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(97)01026-0
Subject(s) - puromycin , in vitro , ribosome , protein biosynthesis , messenger rna , biology , translation (biology) , virus , microbiology and biotechnology , chemistry , rna , biochemistry , genetics , gene
Adequate means for genotype assignment to phenotype is essential in evolutionary molecular engineering. In this study, construction of ‘in vitro virus’ was carried out in which a genotype molecule (mRNA) covalently binds to the phenotype molecule (protein) through puromycin on the ribosome in a cell‐free translation system. Bonding efficiency was ∼10%, thus indicating a population of the in vitro virus to have ∼10 12 protein variants, this number being 10 4 that in the phage display. The in vitro virus is useful for examining protein evolution in a test tube and the results may possibly serve as basis for a general method for selecting proteins possessing the most desirable functions.