Glutamine synthetase gene of Bacillus subtilis |
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Authors: | S H Fisher M S Rosenkrantz A L Sonenshein |
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Affiliation: | Department of Molecular Biology and Microbiology, Tufts University Schools of Medicine, Veterinary Medicine, and Dental Medicine and Sackler School of Graduate Biomedical Sciences, 136 Harrison Avenue, Boston, MA 02111 U.S.A. Tel. (617) 956-6761 |
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Abstract: | The glutamine synthetase gene (glnA) of Bacillus subtilis was purified from a library of B. subtilis DNA cloned in phage lambda. By mapping the locations of previously identified mutations in the glnA locus it was possible to correlate the genetic and physical maps. Mutations known to affect expression of the glnA gene and other genes were mapped within the coding region for glutamine synthetase, as determined by measuring the sizes of truncated, immunologically cross-reacting polypeptides coded for by various sub-cloned regions of the glnA gene. When the entire B. subtilis glnA gene was present on a plasmid it was capable of directing synthesis in Escherichia coli of B. subtilis glutamine synthetase as judged by enzymatic activity, antigenicity, and ability to allow growth of a glutamine auxotroph. By use of the cloned B. subtilis glnA gene as a hybridization probe, it was shown that the known variability of glutamine synthetase specific activity during growth in various nitrogen sources is fully accounted for by changes in glnA mRNA levels. |
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Keywords: | Recombinant DNA nitrogen regulation cloning auxotrophs bacteriophage λ Charon 4A vector S1 nuclease mapping bp base pair BSA bovine serum albumin EtBr ethidium bromide GS glutamine synthetase kb 1000 bp SDS sodium dodecyl sulfate SSC 0.15 MNaCl [] indicates plasmid-carrier state |
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