Open AccessDirect selection of recombinant plasmids with chlorate
Author(s) -
Reiss Jochen,
Klingmüller Walter
Publication year - 1987
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1987.tb02123.x
Subject(s) - plasmid , chlorate , recombinant dna , transformation (genetics) , biology , escherichia coli , cloning (programming) , molecular cloning , cloning vector , mutant , microbiology and biotechnology , multiple cloning site , genetics , vector (molecular biology) , dna , gene , chemistry , peptide sequence , inorganic chemistry , computer science , programming language
Abstract A pair of small plasmid vectors have been constructed, which complement chlorate‐resistant chlB mutants of Escherichia coli to chlorate reduction under anaerobic conditions. This potentially lethal function can be inactivated by the insertion of DNA fragments into a multiple cloning site. After transformation of competent mutant cells with a ligation mixture, simultaneous selection for the plasmid‐coded drug resistance and for the inactivation of the complementing gene directly yields colonies, which harbor recombinant plasmids. It is demonstrated that this cloning system can be used with an efficiency beyond 90%. Vector sequences are provided.