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Cloning of genes for proline and leucine biosynthesis from Brucella abortus by functional complementation in Escherichia coli

Essenberg, R.C.; Sharma, Y.K.

Journal of General Microbiology 139(1): 87-93

1993

ISSN/ISBN: 0022-1287
PMID: 8450311
DOI: 10.1099/00221287-139-1-87
Accession: 008334890
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By selecting for growth of Escherichia coli mutant strains in the absence of the required amino acid, clones were found in a Brucella abortus library carrying genes for glutamyl phosphate reductase (proA) and beta-isopropylmalate dehydrogenase (leuB). These clones hybridized to unique fragments in a genomic digest of B. abortus DNA. The proA-complementing DNA was found in a region of 1.3 kb, which directed the synthesis of a protein of 48,000 Da with a pI of 6.3 in maxicells. The leuB-complementing activity was in a region of 1.4 kb and directed synthesis of a protein of 46,000 Da with a pI of 5.9.

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