Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Cloning and Expression of hpaA Gene of Korean Strain Helicobacter pylori K51 in Oral Vaccine Delivery Vehicle Lactococcus lactis subsp. lactis MG1363

  • Kim Su-Jung (Laboratory of Immunobiology, Department of Microbiology, College of Natural Sciences, Kyungpook National University, Department of Clinical Pathology, Daegu Health College) ;
  • Jun Do-Youn (Institute of Genetic Engineering, Kyungpook National University) ;
  • Yang Chae-Ha (Department of Oriental Medicine, College of Oriental Medicine, Daegu Haany University) ;
  • Kim Young-Ho (Laboratory of Immunobiology, Department of Microbiology, College of Natural Sciences, Kyungpook National University)
  • Published : 2006.02.01
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Abstract

In order to develop an oral vaccine to prevent H. pylori infection, we have expressed the hpaA gene of H. pylori K51 isolated from Korean patients, encoding 29-kDa HpaA that is known to be localized on the cell surface and flagella sheath, in a live delivery vector system, Lactococcus lactis. The hpaA gene, amplified by PCR using the genomic DNA of H. pylori K51, was cloned in the pGEX-2T vector, and the DNA sequence analysis revealed that the hpaA gene of H. pylori K51 had 99.7% and 94.8% identity with individual hpaA genes of the H. pylori 26695 strain (U.K) and the J99 strain (U.S.A). A polyclonal anti-HpaA antibody was raised in rats using GST-HpaA fusion protein as the antigen. The hpaA gene was inserted in an E. coli-L. lactis-shuttle vector (pMG36e) to express in L. lactis. Western blot analysis showed that the expression level of HpaA in the L. lactis transformant remained constant from the exponential phase to the stationary phase, without extracelluar secretion. These results indicate that the HpaA of H. pylori K51 was successfully expressed in L. lactis, and suggest that the recombinant L. lactis expressing HpaA may be applicable as an oral vaccine to induce a protective immune response against H. pylori.

Keywords

References

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