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PMID: 7568014 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The urease locus of Mycobacterium tuberculosis and its utilization for the demonstration of allelic exchange in Mycobacterium bovis bacillus Calmette-Guérin.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 92 ·No. 19 ·1995-09-12 ·Pages 8768-72

Reyrat JM, Berthet FX, Gicquel B

Abstract

The ureABC genes of Mycobacterium tuberculosis were cloned. By using a set of degenerate primers corresponding to a conserved region of the urease enzyme (EC 3.5.1.5), a fragment of the expected size was amplified by PCR and was used to screen a M. tuberculosis cosmid library. Three open reading frames with extensive similarity to the urease genes from other organisms were found. The locus was mapped on the chromosome, using an ordered M. tuberculosis cosmid library. A suicide vector containing a ureC gene disrupted by a kanamycin marker (aph) was used to construct a urease-negative Mycobacterium bovis bacillus Calmette-Guérin mutant by allelic exchange involving replacement of the ureC gene with the aph::ureC construct. To our knowledge, allelic exchange has not been reported previously in the slow-growing mycobacteria. Homologous recombination will be an invaluable genetic tool for deciphering the mechanisms of tuberculosis pathogenesis, a disease that causes 3 x 10(6) deaths a year worldwide.

Related Genes
ureA ureB ureC
MeSH Terms
Alleles Amino Acid Sequence Base Sequence Blotting, Southern Chromosome Mapping Genome, Bacterial Genomic Library Genotype Molecular Sequence Data Mutagenesis, Insertional Mycobacterium bovis/genetics Mycobacterium tuberculosis/enzymology,genetics Polymerase Chain Reaction Recombination, Genetic Sequence Analysis, DNA Sequence Homology, Amino Acid Species Specificity Urease/genetics
Chemicals
Urease
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Reyrat J M
Unité de Génétique Mycobactérienne, Centre National de la Recherche Scientifique, Institut Pasteur 25, Paris, France.
Berthet F X
Gicquel B
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.92.19.8768
Published
1995-09-12
Pages
8768-72
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC41048
Subset
IM
Databases
GENBANK
L41141
External Links
PubMed source DOI: 10.1073/pnas.92.19.8768 PMC Full Text View journal details
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