Facile construction of mutations in Haemophilus ducreyi using lacZ as a counter-selectable marker |
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Authors: | Joel A Bozue Laurie Tarantino Robert S Munson Jr. |
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Affiliation: | Children's Hospital Research Foundation, Room W402, 700 Children's Drive, Columbus, OH 43205, USA;Department of Pediatrics, The Ohio State University, Columbus, OH 43205, USA;Department of Medical Microbiology and Immunology, The Ohio State University, Columbus, OH 43205, USA |
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Abstract: | Haemophilus ducreyi is a Gram-negative bacterium which is the causative agent of chancroid, an ulcerative sexually transmitted disease. In order to understand the pathogenesis of H. ducreyi disease, studies designed to identify potential virulence determinants and construct mutants deficient in the elaboration of these determinants have been undertaken in several laboratories. At the present time, construction of isogenic mutants is accomplished by electroporation of linearized DNA containing insertionally inactivated H. ducreyi genes followed by selection for the resistance marker encoded on the inactivated gene. In our experience, certain mutants are difficult to construct using this procedure. In the construction of strains containing lacZ as a reporter gene, we observed that the growth of lacZ expressing H. ducreyi was inhibited in the presence of X-gal. We have exploited this observation to develop a new strategy for the construction of isogenic H. ducreyi mutants. |
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Keywords: | Haemophilus ducreyi Isogenic mutant construction LacZ |
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