Open AccessCloning of the structural gene (ompA) for an integral outer membrane protein of Escherichia coli K-12.
Author(s) -
Ulf Henning,
Hans-Dieter Royer,
R. M. Teather,
Ingrid Hindennach,
Cornelis P. Hollenberg
Publication year - 1979
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.76.9.4360
Subject(s) - bacterial outer membrane , escherichia coli , microbiology and biotechnology , biology , lytic cycle , ecori , plasmid , vesicle associated membrane protein 8 , hspa2 , bacteriophage , membrane protein , gene , peptide sequence , biochemistry , virology , membrane , virus
The gene (ompA) for the major outer membrane protein II* from Escherichia coli K-12 has been cloned on a 5-megadalton EcoRI fragment by using phage lambda as vector. The gene is expressed during the lytic cycle of the recombinant phage and the insoluble membrane-bound protein was detected in phage plaques with a simple radioimmunoassay. Transfer of the EcoRI fragment into plasmid pSC101 and expression in a host lacking protein II* led to overproduction of protein II* and decreased production of two other major outer membrane proteins. Expression of the plasmid pSC101-ompA+ in minicells derived from an ompA minicell-producing strain led to synthesis, at high rates, of this protein and massive accumulation of a second cell envelope protein most likely representing the biosynthetic precursor of protein II*.