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cDNA–Protein Fusions: Covalent Protein–Gene Conjugates for the In Vitro Selection of Peptides and Proteins
Author(s) -
Kurz Markus,
Gu Ke,
AlGawari Amal,
Lohse Peter A.
Publication year - 2001
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/1439-7633(20010903)2:9<666::aid-cbic666>3.0.co;2-#
Subject(s) - complementary dna , in vitro , biochemistry , conjugate , gene , chemistry , biology , covalent bond , organic chemistry , mathematical analysis , mathematics
We report a method for the synthesis of covalent cDNA‐protein fusions for protein display applications. A branched mRNA template was developed which carries a peptidyl acceptor and a reverse transcription primer at the 3′‐end. Translation in vitro followed by reverse transcription produced a protein covalently bonded to its encoding cDNA. Both single‐ and double‐stranded cDNA–protein fusions were prepared. cDNA–protein fusions are stable in alkali and resistant to ribonucleases. Their simple preparation and their resistance towards degradation should make cDNA–protein fusions a useful tool for the in vitro selection and evolution of high affinity ligands from large libraries of polypeptides.