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Operon fusions of mu d1 ap lac to the l proline biosynthetic genes of escherichia coli k 12

Operon fusions of mu d1 ap lac to the l proline biosynthetic genes of escherichia coli k 12

Current Microbiology 18(2): 113-118

ISSN/ISBN: 0343-8651

Operon fusions to the promoter of either the proA, proB, or proC genes of the proline biosynthetic pathway were obtained by the use of the Mu d1(Ap, lac) bacteriophage. These fusions were further stabilized by transformation with plasmid pGW600 containing the wild-type Mu repressor gene or by transduction with phage .lambda.pSG1. The level of .beta.-galactosidase in the fusion strains was not affected by the persence of exogenously added L-proline or high concentrations of NaCl in the growth medium. A Tn5 insertion near proBA increased .beta.-galacosidase expression 140- to 200-fold in strains carrying the proA-lac and proB-lac fusions, but the level of this enzyme was unaltered in strains carrying the proC-lac fusion. The Tn5 insertion increased intracellular proline concentrations 8- to 10-fold, suggesting that mechanisms other than allosteric inhibition may regulate proline biosynthesis, but did not confer osmotolerance to cells growing in a medium with a high concentration of salt.

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Accession: 007618441

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