Identification of mycobacterium tuberculosis DNA sequences encoding exported proteins by using phoA gene fusions
| Autor: | B Gicquel, J. Timm, D. Portnoi, J. Rauzier, A. Murray, Eng Mong Lim, G. Torrea |
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| Rok vydání: | 1995 |
| Předmět: |
DNA
Bacterial Recombinant Fusion Proteins Genetic Vectors Molecular Sequence Data Protein Sorting Signals Microbiology beta-Lactamases Mixed Function Oxygenases Mycobacterium Mycobacterium tuberculosis Bacterial Proteins Genomic library Amino Acid Sequence Molecular Biology Peptide sequence Mycobacterium leprae Gene Genetics Genomic Library biology Base Sequence Biological Transport biology.organism_classification Alkaline Phosphatase Genes Bacterial bacteria Mycobacterium fortuitum Research Article |
| Zdroj: | Journal of bacteriology. 177(1) |
| ISSN: | 0021-9193 |
| Popis: | The activity of bacterial alkaline phosphatase (PhoA) is dependent on it being exported across the plasma membrane. A plasmid vector (pJEM11) allowing fusions between phoA and genes encoding exported proteins was constructed to study protein export in mycobacteria. Introduction of the Mycobacterium fortuitum beta-lactamase gene (blaF*) into this vector led to the production in M. smegmatis of protein fusions with PhoA activity. A genomic library from M. tuberculosis was constructed in pJEM11 and screened in M. smegmatis for clones with PhoA activity. Sequences of the M. tuberculosis inserts directing the production of protein fusions in these PhoA-positive clones were determined. They include part of the already-known exported 19-kDa lipoprotein, a sequence with similarities to the exported 28-kDa antigen from M. leprae, a sequence encoding a protein sharing conserved amino acid motifs with stearoyl-acyl-carrier-protein desaturases, and unknown sequences. This approach thus appears to identify sequences directing protein export, and we expect that more extensive screening of such libraries will lead to a better understanding of protein export in M. tuberculosis. |
| Databáze: | OpenAIRE |
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