Single-step PCR amplification and enzyme restriction analysis of the entire Helicobacter pylori cytotoxin vacA gene for genetic variability studies

Abstract To monitor changes along the entire Helicobacter pylori vacA gene we carried out full-length single-step PCR amplification in 21 gastritis and gastric cancer isolates. HindIII restriction analysis led us to detect a >400-bp internal insertion in vacA subsequently shown to be a direct 451...

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Bibliographic Details
Published in:FEMS microbiology letters 1999-09, Vol.178 (1), p.55-62
Main Authors: Perales, G., Sanchez, J., Mohar, A., Lara-Lemus, R., Hernández, A., Herrera-Goepfert, R., Barrios-Jacobo, I., Ayala, G.
Format: Article
Language:English
Subjects:
Amplification
Bacterial Proteins - genetics
Bacteriology
Biological and medical sciences
Fundamental and applied biological sciences. Psychology
Gastric cancer
Gastritis
Gene duplication
Genes
Genes, Bacterial
Genetic variability
Genetic Variation
Genetics
Helicobacter pylori
Helicobacter pylori - chemistry
Helicobacter pylori - genetics
Humans
Insertion
Internal genetic duplication
Microbiology
Non‐cytotoxic Helicobacter pylori
PCR amplification of vacA gene
Polymerase Chain Reaction - methods
Restriction Mapping - methods
vacA gene diversity
VacA protein
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Summary:Abstract To monitor changes along the entire Helicobacter pylori vacA gene we carried out full-length single-step PCR amplification in 21 gastritis and gastric cancer isolates. HindIII restriction analysis led us to detect a >400-bp internal insertion in vacA subsequently shown to be a direct 451-bp gene duplication. We found HindIII profiles for 16 genes that allowed their grouping into two restriction patterns that were related to theoretical profiles for previously sequenced Western genes. Comparisons with theoretical HindIII patterns for Japanese isolates appear suggestive of geographical H. pylori clonality. Full-length single-step PCR amplification seems suitable for quick restriction pattern assignment and detection of gene size changes.
ISSN:0378-1097
1574-6968
DOI:10.1111/j.1574-6968.1999.tb13759.x