Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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IVET-based Identification of Virulence Factors in Vibrio vulnificus MO6-24/O

  • Lee, Ko-Eun (Department of Life Science and Interdisciplinary Program of Integrated Biotechnology, Sogang University) ;
  • Bang, Ji-Sun (Department of Life Science and Interdisciplinary Program of Integrated Biotechnology, Sogang University) ;
  • Baek, Chang-Ho (Department of Life Science and Interdisciplinary Program of Integrated Biotechnology, Sogang University) ;
  • Park, Dae-Kyun (Department of Life Science and Interdisciplinary Program of Integrated Biotechnology, Sogang University) ;
  • Hwang, Won (Department of Life Science and Interdisciplinary Program of Integrated Biotechnology, Sogang University) ;
  • Choi, Sang-Ho (Department of Food Science and Technology, Seoul National University) ;
  • Kim, Kum-Soo (Department of Life Science and Interdisciplinary Program of Integrated Biotechnology, Sogang University)
  • Published : 2007.02.28
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Abstract

Vibrio vulnificus is an opportunistic pathogen that causes septicemia in humans. To identify the genes associated with its pathogenicity, in vivo expression technology (IVET) was used to select genes specifically expressed in a host, yet not significantly in vitro. Random lacZ-fusions in the genome of V vulnificus strain MO6-24/O were constructed using an IVET vector, pSG3, which is a suicide vector containing promoterless-aph and -lacZ as reporter genes. A total of ${\sim}18,000$ resulting library clones were then intraperitoneally injected into BALB/c mice using a colony forming unit (CFU) of $1.6{\times}10^6$. Two hours after infection, kanamycin was administered at $200{mu}g$ per gram of mouse weight. After two selection cycles, 11 genes were eventually isolated, which were expressed only in the host. Among these genes, VV20781 and VV21007 exhibiting a homology to a hemagglutinin gene and tolC, respectively, were selected based on having the highest frequency. When compared to wild-type cells, mutants with lesions in these genes showed no difference in the rate of growth rate, yet a significant decrease in cytotoxicity and the capability to form a biofilm.

Keywords

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