Abstract
The genus Helicobacter currently comprises over 15 different species with members described as microaerophilic gram-negative spiral rods with a G + C content of 35–38%. The most medically important Helicobacter is Helicobacter pylori, a human pathogen that has been causally linked to the development of gastritis, gastric ulcers, and gastric cancer (1,2). The need to fulfill, at the molecular level, Koch’s Postulates for any microbial pathogen investigated demands that the organism be applicable to all the molecular genetic techniques available. Ultimately one must be able to prepare DNA from the organism, clone and manipulate the DNA in the laboratory, and most importantly, insert DNA into the isolate. Only in this manner can isogenic mutants be created, allowing for the proper comparison of wild-type and mutant in an appropriate model to determine potential virulence factors.
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References
Parsonnet, J., Friedman, G. D., Vandersteen, D. P., Chang, Y., Vogelman, J. H., Orentreich, N., and Sibley, R. K. (1991) Helicobacter pylori infection and the risk of gastric carcinoma. New Engl. J. Med. 325, 1127–1131.
Nomura, A., Stemmerman, G. N., Chyou, P. H., Kato, I., Perez-Perez, G. I., and Blaser, M. (1991) Helicobacter pylori infection and gastric carcinoma among Japanese. New Engl. J. Med. 325, 1132–1136.
Miller, J., Dower, W. J., and Tompkins, L. S. (1988) High-voltage electroporation of bacteria: genetic transformation of C. jejuni with plasmid DNA. Proc. Natl. Acad. Sci. USA 85, 856–860.
Segal, E. D., and Tompkins, L. S. (1993) Transformation of Helicobacter pylori by electroporation. Biotechniques 14, 225,226.
Tsuda, M., Karita, M., and Nakazawa, T. (1993) Genetic transformation in H. pylori. Microbiol. Immunol. 37, 85–89.
Nedenskov-Sorensen, P., Bukholm, G., and Bovre, K. (1990) Natural competence for genetic transformation in Campylobacter pylori. J. Infect. Dis. 161, 365.
Ferrero, R. L., Cussac, V., Courcoux, P., and Labigne, A. (1992) Construction of isogenic urease-negative mutants of Helicobacter pylori by allelic exchange. J. Bacteriol. 174, 4212–4217.
Taylor, D. E. (1992) Genetics of Campylobacter and Helicobacter. Ann. Rev. Microbiol. 46, 35–64.
Taylor, D. E., Roos, K. P., and Wang, Y. (1993) Transformation of H. pylori by chromosomal metronidazole resistance and by a plasmid with a chloramphenicol resistance marker, in Campylobacter Meets Helicobacter: The Joint Meeting. Brussels. Acta. Gastroenterol. Belg. 56(Suppl.), 106.
Haas, R., Meyer, T. F., and van Putten, J. P. M. (1993) Aflagellated mutants of H. pylori generated by genetic transformation of naturally competent straits using transposon shuttle mutagenesis. Molecular Microbiol. 8, 753–760.
Suerbaum, S., Josenhans, C, and Labigne, A. (1993) Cloning and genetic characterization of the H. pylori and H. mustelae flaB flagellin genes and construction of H. pylori flaA-and flaB-negative mutants by electroporation-mediated allelic exchange. J. Bacteriol. 175, 3278–3288.
Labigne-Roussel, A., Harel, J., and Tompkins, L. S. (1987) Gene transfer from E. coli to Campylobacter species: development of shuttle vectors for genetic analysis of C. jejuni. J. Bacteriol. 169, 5320–5323.
Labigne, A., Courcoux, P., and Tompkins, L. (1992) Cloning of C. jejuni genes required for leucine biosynthesis, and construction of leu negative mutant of C. jejuni by shuttle transposon mutagenesis. Res. Microbiol. 143, 15–26.
Haas, R., Odenbreit, S., Heuermann, D., and Schmitt, W. (1993) Transposon shuttle mutagenesis as a general tool to identify and genetically define virulence determinants of H. pylori, in Campylobacter Meets Helicobacter: The Joint Meeting. Brussels. Acta. Gastroenterol. Belg. 56(Suppl.), 103.
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© 1995 Humana Press Inc.
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Segal, E.D. (1995). Electroporation of Helicobacter pylori . In: Nickoloff, J.A. (eds) Electroporation Protocols for Microorganisms. Methods in Molecular Biology™, vol 47. Humana Press. https://doi.org/10.1385/0-89603-310-4:179
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DOI: https://doi.org/10.1385/0-89603-310-4:179
Publisher Name: Humana Press
Print ISBN: 978-0-89603-310-8
Online ISBN: 978-1-59259-534-1
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