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Complementation system for Helicobacter pylori.

 Jinmoon Kim  ;  Sung-Whan Kim  ;  Sungil Jang  ;  D. Scott Merrell  ;  Jeong-Heon Cha 
 JOURNAL OF MICROBIOLOGY, Vol.49(3) : 481-486, 2011 
Journal Title
Issue Date
Bacterial Proteins/chemistry ; Bacterial Proteins/genetics ; Base Sequence ; Chromosomes, Bacterial/genetics ; Culture Media ; DNA, Bacterial/analysis ; DNA, Bacterial/genetics ; DNA, Intergenic/genetics* ; GeneticComplementationTest* ; Helicobacter pylori/classification ; Helicobacter pylori/genetics* ; Molecular Sequence Data ; Sequence Analysis, DNA ; Transformation, Bacterial*
H. pylori ; intergenic region ; complementation system
Previously Langford et al. (2006) developed the pIR203C04 complementation system for Helicobacter pylori, which can be used to complement and restore phenotypic effects in H. pylori mutant, and furthermore they used the complementation system in vivo experiments to animals without altering the ability of strain SSI to colonize mice. In their previous study, the pIR203C04 was able to transform 26695, SSI, J99, and 43504 H. pylori strains by an electroporation method. However, in the present study using a natural transformation the pIR203C04 transformed only 26695 H. pylori but not SSI, J99, 7.13, and G27 H. pylori strains. Since the useful complementation system has a limitation of narrow selection among H. pylori strains, we redesigned the complementation system for the improvement. The same intergenic chromosomal site between hp0203 and hp0204 was utilized for the new complementation system because the insertion at the intergenic site didn't show any polar effects and disruption of other H. pylori genes. The genome sequence analysis showed that the intergenic regions among H. pylori strains may have too low homology to each others to do a homologous recombination. Thus, in addition to the short intergenic region, the fragments of the new complementation system included 3' conserved parts of hp0203 and hp0204 coding regions. Between the fragments there are a chloramphenicol acetyltransferase cassette and multicloning sites, resulting in pKJMSH. DNA fragment of the interest can be cloned into the multicloning sites of pKJMSH and the fragment can be integrated at the intergenic region of H. pylori chromosome by the homologous recombination. Indeed, by the natural transformation, pKJMSH was able to transform all five H. pylori strains of 26695, SSI, J99, 7.13, and G27, which are common for the investigation of molecular pathogenesis. Thus, the new pKJMSH complementation system is applicable to most H. pylori wild-type stains.
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2. College of Dentistry (치과대학) > Research Institute (부설연구소) > 1. Journal Papers
2. College of Dentistry (치과대학) > Dept. of Oral Biology (구강생물학교실) > 1. Journal Papers
Yonsei Authors
Kim, Sung Whan(김성환)
Kim, Jin Moon(김진문)
Jang, Sungil(장성일) ORCID logo https://orcid.org/0000-0001-6144-6899
Cha, Jeong Heon(차정헌) ORCID logo https://orcid.org/0000-0002-9385-2653
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