期刊
MOLECULAR MICROBIOLOGY
卷 37, 期 5, 页码 1052-1065出版社
WILEY
DOI: 10.1046/j.1365-2958.2000.02049.x
关键词
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资金
- NIDDK NIH HHS [R01DK53707] Funding Source: Medline
Helicobacter pylori cells are naturally competent for the uptake of both plasmid and chromosomal DNA. However, we demonstrate that there are strong barriers to transformation of H. pylori strains by plasmids derived from unrelated strains. We sought to determine the molecular mechanisms underlying these barriers. Transformation efficiency was assessed using pHP1, an Escherichia coli-H. pylori shuttle vector conferring kanamycin resistance. Transformation of 33 H. pylori strains was attempted with pHP1 purified from either E. coli or H. pylori, and was successfully introduced into only 11 strains. Digestion of H. pylori chromosomes with different restriction endonucleases (REs) showed that DNA methylation patterns vary substantially among strains. The strain most easily transformed, JP26, was found to have extremely low endogenous RE activity and to lack a restriction-modification (R-M) system, homologous to Mbol, which is highly conserved among H. pylori strains. When we introduced this system to JP26, pHP1 from Mbol.M- JP26, but not from wild-type JP26, transformed Mbol R-M+ JP26 and heterologous Mbol R-M+ wild-type H. pylori strains. Parallel studies with pHP1 from dam(+) and dam(-) E. coli strains confirmed these findings. These data indicate that the endogenous REs of H. pylori strains represent a critical barrier to interstrain plasmid transfer among H. pylori.
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