Expression and Purification of Glutamine Synthetase Cloned from Bacteroides fragilis
- 1 October 1986
- journal article
- research article
- Published by Microbiology Society in Microbiology
- Vol. 132 (10) , 2827-2835
- https://doi.org/10.1099/00221287-132-10-2827
Abstract
A glutamine synthetase (GS) gene, glnA, from Bacteroides fragilis was cloned on a recombinant plasmid pJS139 which enabled Escherichia coli glnA deletion mutants to utilize (NH4)2SO4 as a sole source of nitrogen. DNA homology was not detected between the B. fragilis glnA gene and the E. coli glnA gene. The cloned B. fragilis glnA gene was expressed from its own promoter and was subject to nitrogen repression in E. coli, but it was not able to activate histidase activity in an E. coli glnA ntrB ntrC deletion mutant containing the Klebsiella aerogenes hut operon. The GS produced by pJS139 in E. coli was purified: it had an apparent subunit Mr of approximately 75,000, which is larger than that of any other known bacterial GS. There was very slight antigenic cross-reactivity between antibodies to the purified cloned B. fragilis GS and GS subunit of wild-type E. coli.Keywords
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