PMID: 9432340Feb 12, 1998Paper

Rapid polymerase chain reaction screening of Helicobacter pylori chromosomal point mutations

Helicobacter
Z Ge, D E Taylor

Abstract

Microdiversity (within individual genes) in the genomes of different Helicobacter pylori strains has been demonstrated to be more frequent than that seen in other prokaryotes. Point mutations in some genes, such as the vacA and 23S ribosomal RNA genes could result in the alteration of pathogenicity or antibiotic susceptibility of individual H. pylori strains. Development of a simple, rapid, and reliable screening method would be useful in the molecular characterization of genetic variation among different H. pylori strains. The copP gene from H. pylori UA802 was used as a model for developing a mutation screening method. Four point mutations were introduced into the copP gene by in vitro site-directed mutagenesis and were verified by DNA sequencing. The mutated copP gene replaced the wild-type locus by natural transformation and homologous recombination. The site-specific mutants were screened by polymerase chain reaction (PCR) using 3'-end mismatched primers. The origins of the PCR fragments were demonstrated by Southern hybridization with the copP-derived DNA probe. Three of these four mutations were characterized by PCR with the specific primers that contained the 3'-terminal nucleotide complementary only to the mutated nucl...Continue Reading

References

Nov 24, 1979·Nucleic Acids Research·H C Birnboim, J Doly
Aug 15, 1996·FEMS Microbiology Letters·Y J Debets-OssenkoppC M Vandenbroucke-Grauls
Feb 1, 1996·Antimicrobial Agents and Chemotherapy·J VersalovicM F Go

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Citations

Nov 9, 2011·Saudi Journal of Gastroenterology : Official Journal of the Saudi Gastroenterology Association·Amin T AbadiAshraf M Mobarez
Sep 1, 2015·Gastroenterology Clinics of North America·Francis MégraudPhilippe Lehours
Apr 17, 2012·La Revue de médecine interne·J-D de Korwin
Jun 8, 2002·Expert Review of Molecular Diagnostics·Sandra HjalmarssonLars Engstrand
Feb 28, 2002·The American Journal of Gastroenterology·Paul R HarrisPhillip D Smith
Feb 4, 2005·APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica·Joanne L Simala-Grant, Diane E Taylor
Nov 18, 1998·Journal of Clinical Microbiology·N MisawaM J Blaser

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