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An antibody VH domain with a lox ‐Cre site integrated into its coding region: bacterial recombination within a single polypeptide chain
Author(s) -
Davies Julian,
Riechmann Lutz
Publication year - 1995
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 - 10.1016/0014-5793(95)01313-x
Subject(s) - cre lox recombination , recombination , microbiology and biotechnology , plasmid , mutant , phage display , coding region , immunoglobulin light chain , phagemid , gene , biology , genetics , chemistry , antibody , bacteriophage , escherichia coli , transgene , genetically modified mouse
Bacterial lox ‐Cre recombination within a single antibody VH domain was achieved through integration of a loxP site into its coding sequence. The 5′ half of the VH gene, in which the H2 loop was replaced by a mutant loxP site, was fused to geneIII in an ‘acceptor’ fd‐phage vector containing also a wild type loxP site. With a ‘donor’ plasmid vector harbouring the 3′ half of the VH gene flanked by the same, differing loxP sites it recombined into a full‐length VH with the loxP site‐H2 loop. This VH was purified from bacterial periplasm, where it folded into a typical immunoglobulin domain. The system allows the generation of large VH repertoires using lox ‐Cre recombination.