Use of lambda unc transducing bacteriophages in genetic and biochemical characterization of H super(+)-ATPase mutants of Escherichia coli

The eight subunits of the H super(+)-ATPase of Escherichia coli) are coded by the genes of the unc operon, which maps between bglB and ansA. A collection of unc mutations were transferred via P1 transduction into a strain in which lambda c1857 S7 was inserted into bglB. The lambda phage was induced,...

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Bibliographic Details
Published in:Journal of bacteriology 1983-01, Vol.156 (3), p.1078-1092
Main Authors: Mosher, ME, Peters, L K, Fillingame, R H
Format: Article
Language:English
Subjects:
adenosinetriphosphatase
Escherichia coli
mutants
operons
phage lambda
transduction
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Summary:The eight subunits of the H super(+)-ATPase of Escherichia coli) are coded by the genes of the unc operon, which maps between bglB and ansA. A collection of unc mutations were transferred via P1 transduction into a strain in which lambda c1857 S7 was inserted into bglB. The lambda phage was induced, and asnA super(+) transducing phage that carried unc) were selected. Transducing phage carrying mutations in the uncA , B, D, E, and E genes were used for complementation analysis with a collection of unc mutants, including mutants which had been reported previously but not genetically characterized. Two mutants (uncE105 and uncE107) altered in the proteolipid (omega) subunit of F sub(0) were not complemented by any of the lambda phage, even though both mutants had a fully functional F sub(1) ATPase and therefore normal A and D genes. The lambda phage proved to essential in characterizing several mutants defective in F sub(o)-mediated H super(+) translocation. The unc) gene products were overproduced by heat induction of the lysogenized lambda unc phage to determine whether all of F sub(0) subunits were in the membrane.
ISSN:0021-9193