Identification and genetic characterization of the Pseudomonas aeruginosaleuB gene encoding 3-isopropylmalate dehydrogenase |
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Authors: | T T Hoang and H P Schweizer |
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Institution: | (1) Department of Microbiology, Colorado State University, Fort Collins, CO 80523, USA, US |
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Abstract: | The Pseudomonas aeruginosa leuB gene, encoding 3-isopropylmalate dehydrogenase, was identified upstream of asd, encoding aspartate-β-semialdehyde dehydrogenase. Genetic analysis indicated that leuB is identical to the previously mapped gene defined by the leu-10 allele. The chromosomal leuB locus was inactivated by gene replacement. The insertions had no adverse effect on expression of the downstream asd gene but resulted in leucine auxotrophy. The leuB gene encodes a protein containing 360 amino acids (with a molecular weight of 39153), which was expressed in Escherichia coli as a M, 42000 protein. The results suggested that, in contrast to the situation in other bacteria (E. coli, Salmonella typhimurium and Bacillus subtilis) the P. aeruginosa leuB gene is physically separated from the genes encoding the other enzymes of the isopropylmalate pathway.
Received: 15 August 1996 / Accepted: 23 October 1996 |
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Keywords: | Pseudomonas aeruginosa Leucine biosynthesis 3-Isopropylmalate dehydrogenase Cloning Genetic analysis |
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