Journal
MUTATION RESEARCH-GENETIC TOXICOLOGY AND ENVIRONMENTAL MUTAGENESIS
Volume 535, Issue 2, Pages 141-145Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/S1383-5718(02)00292-9
Keywords
H. pylori; rpsL; mutation; streptomycin-resistance
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Several studies have revealed that the Helicobacter pylori genome differs markedly from strain to strain, perhaps as a result of mutations arising during persistent infection and/or related to the observed variation in virulence. The development of a detection system for mutations in H. pylori genes might therefore help us to develop a better understanding of its mutability, and in this way help us to develop plans for investigating the relationship between its genomic variability and the pathogenesis of various gastric and duodenal diseases associated with the long-term H. pylori infections. We have therefore begun a study of H. pylori mutability using the endogenous rpsL gene as a marker. Spontaneous mutant frequencies were measured and compared among H. pylori strains, after incubation on plates containing 50 mug/ml of streptomycin for 10 days as a selection procedure. The rpsL gene of each streptomycin-resistant (Str(r)) mutant was amplified by polymerise-chain-reaction (PCR) and sequenced. All of the mutations we characterized were localized at codons 43 or 88 of the rpsL gene and were base transitions from A to G, replacing lysine with arginine. This is in contrast to the spontaneous Strr mutants isolated from Escherichia coli, which resulted from either A to G transitions at lysine codons 42 and 87, or A to T or C transversions at lysine codon 42. The spontaneous mutant frequencies of the rpsL gene in H. pylori were of the order of 10(-9), and there were significant differences in spontaneous mutant frequencies among the strains tested. This mutation detection system might be of value in screening clinical isolates for H. pylori mutator phenotypes. (C) 2002 Elsevier Science B.V. All rights reserved.
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