Expression cloning of cDNA by phage display selection
Author(s) -
Janice Light
Publication year - 1996
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/24.21.4367
Subject(s) - biology , complementary dna , phage display , cloning (programming) , leucine zipper , cdna library , genomic library , microbiology and biotechnology , phagemid , expression cloning , molecular cloning , selection (genetic algorithm) , bacteriophage , genetics , peptide sequence , gene , escherichia coli , antibody , artificial intelligence , computer science , programming language
Expression cloning of a mouse kappa chain fragment has been achieved from a cDNA library by display of expressed proteins on filamentous phage and affinity selection for binding to anti-mouse Fab antibodies. Expressed proteins were anchored to the phage coat by a synthetic, anti-parallel leucine zipper, which had been selected from a semi-randomized zipper library for the ability to connect a test protein to phage. From a library of 4 x 10(6) transformants, two separate clones displaying different size cDNA inserts were recovered after four selection rounds. These results further demonstrate the utility of phage display for cDNA expression cloning.
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