Efficient allelic exchange and transposon mutagenesis in Mycobacterium avium by specialized transduction Journal Article
| Authors: | Otero, J.; Jacobs, W. R. Jr; Glickman, M. S. |
| Article Title: | Efficient allelic exchange and transposon mutagenesis in Mycobacterium avium by specialized transduction |
| Abstract: | Mycobacterium tuberculosis and Mycobacterium avium are pathogenic slow-growing mycobacteria that cause distinct human diseases. In contrast to recent advances in M. tuberculosis genetics and pathogenesis investigation, M. avium has remained genetically intractable and, consequently, its pathogenic strategies remain poorly understood. Here we report the successful development of efficient allelic exchange and transposon mutagenesis in an opaque clinical strain of M. avium by specialized transduction. Efforts to disrupt the leuD gene of M. avium by specialized transduction were successful but were complicated by inefficient isolation of recombinants secondary to high spontaneous antibiotic resistance. However, by using this leucine auxotroph as a genetic host and the Streptomyces coelicolor leuD gene as a selectable marker, we achieved efficient allelic exchange at the M. avium pcaA locus. A leud-marked transposon delivered by specialized transduction mutagenized M. avium with efficiencies similar to M. tuberculosis. These results establish a system for random and directed mutagenesis of M. avium. In combination with the forthcoming M. avium genome sequence, these tools will allow the distinct physiologic and pathogenic properties of M. avium to be dissected in molecular detail. |
| Keywords: | gene mutation; gene sequence; pathogenesis; nonhuman; allele; genes; gene locus; alleles; pathology; bacterial strain; bacteria (microorganisms); transduction, genetic; antibiotic resistance; mycobacterium tuberculosis; bacterial proteins; gene disruption; microbiology; site directed mutagenesis; mycobacterium; corynebacterineae; mutagenesis; diseases; genetic markers; transposon; mutagenesis, insertional; bacterial gene; genetic complementation test; bacteria; streptomyces; prokaryota; actinobacteria (class); mycobacterium avium; article; transposons; insertion sequences; uncultured actinomycete; bacterium transduction; leud gene; streptomyces coelicolor; hydro-lyases |
| Journal Title: | Applied and Environmental Microbiology |
| Volume: | 69 |
| Issue: | 9 |
| ISSN: | 0099-2240 |
| Publisher: | American Society for Microbiology |
| Date Published: | 2003-09-01 |
| Start Page: | 5039 |
| End Page: | 5044 |
| Language: | English |
| DOI: | 10.1128/aem.69.9.5039-5044.2003 |
| PUBMED: | 12957884 |
| PROVIDER: | scopus |
| PMCID: | PMC194949 |
| DOI/URL: | |
| Notes: | Export Date: 12 September 2014 -- Source: Scopus |
