protein-dna complexes in mycobacteriophage l5 integrative recombination.the temperate mycobacteriophage l5 integrates site specifically into the genomes of mycobacterium smegmatis, mycobacterium tuberculosis, and mycobacterium bovis bacillus calmette-guérin. this integrative recombination event occurs between the phage l5 attp site and the mycobacterial attb site and requires the phage-encoded integrase and mycobacterial-encoded integration host factor mihf. here we show that attp, int-l5, and mihf assemble into a recombinationally active complex, the intasome, whic ...19999882658
identification of three cytotoxic early proteins of mycobacteriophage l5 leading to growth inhibition in mycobacterium smegmatis.mycobacteriophage l5 is a temperate phage with a broad host range among the fast- and slow-growing mycobacteria such as mycobacterium smegmatis, mycobacterium tuberculosis, mycobacterium avium and mycobacterium ulcerans. l5 switches off host protein synthesis during the early stage of lytic growth, as was previously shown by protein expression profiling. also, lethal genetic elements have been identified in l5 based on the fact that transformants could not be obtained with these genes. using an ...200818667563
dna sequence, structure and gene expression of mycobacteriophage l5: a phage system for mycobacterial genetics.genetic studies of mycobacterium tuberculosis and other mycobacterial pathogens have suffered from the lack of a sophisticated genetic system. to address this issue we have developed a viral system through a detailed characterization of mycobacteriophage l5, a temperate phage that infects both fast- and slow-growing mycobacteria. we describe here the complete dna sequence of the l5 genome and initial characterization of l5 virion structure and gene expression. in addition to providing a genetic ...19938459766
efficient switching of mycobacteriophage l5-based integrating plasmids in mycobacterium tuberculosis.we previously used a mycobacteriophage l5-derived integrating vector to demonstrate that glne and arok are essential genes in mycobacterium tuberculosis by showing that we were unable to excise the integrated vector when it carried the only functional copy of these genes. we tested three systems to replace the integrated copy with alternative alleles. the most efficient method was to transform the strain with a second copy of the integrating vector. excision of the resident vector and integratio ...200314680701
expression systems for study of mycobacterial gene regulation and development of recombinant bcg vaccines.successful genetic engineering of mycobacteria is crucial for developing new approaches to combat tuberculosis as well as for dissecting out the molecular basis of pathogenesis of mycobacterium tuberculosis. we have constructed a mycobacterium-escherichia coli shuttle expression vector psd5. it carries a modular expression cassette which provides sites for cloning of promoters, a ribosome binding site (rbs) with an appropriately placed initiation codon and multiple cloning sites for cloning the ...19989618292
mycobacteriophage l5 infection of mycobacterium bovis bcg: implications for phage genetics in the slow-growing mycobacteria.mycobacteriophage l5 is a well-characterized temperate phage that forms stable lysogens in mycobacterium smegmatis. the host range of l5 is, however, unclear because previous reports suggested that it does not infect slow-growing mycobacteria such as mycobacterium tuberculosis and bacille calmette-guerin (bcg). moreover, luciferase reporter phage derivatives of l5 failed to produce light from bcg, suggesting that infection is blocked at or before the stage of dna injection. in this study, we dem ...19979427405
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