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Site‐specific recombination in human cells catalyzed by the wild‐type integrase protein of coliphage HK022
Author(s) -
Kolot Mikhail,
Meroz Adi,
Yagil Ezra
Publication year - 2003
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.10747
Subject(s) - integrase , plasmid , coliphage , integrases , green fluorescent protein , biology , recombination , trans acting , wild type , bacteriophage , microbiology and biotechnology , site specific recombination , gene , chemistry , genetics , escherichia coli , recombinase , mutant
The activity of the Integrase (Int) protein encoded by coliphage HK022 was tested in a human cell culture. Plasmids were constructed as substrates that carry the sites of the integration reaction ( attP and attB ) or the sites of excision ( attL and attR ). The site‐specific recombination reactions were monitored in cis and in trans configurations by the expression of the green fluorescent protein (GFP) as a reporter. Cells cotransfected with the substrate plasmid(s) and with a plasmid that expresses the wild‐type Int show efficient integration as well as excision in both configurations. The wild‐type Int was active in the human cells without the need to supply the accessory proteins integration host factor (IHF) and excisionase (Xis) that are indispensable for the reaction in the bacterial host. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 84: 56–60, 2003.