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Expression of Anthrax Lethal Factor, a Major Virulence Factor of Anthrax, in Saccharomyces cerevisiae  

Hwang Hyehyun (Department of Chemistry, College of Natural Science, Hanyang University)
Kim Joungmok (Department of Chemistry, College of Natural Science, Hanyang University)
Choi Kyoung-Jae (Department of Chemistry, College of Natural Science, Hanyang University)
Chung Hoeil (Department of Chemistry, College of Natural Science, Hanyang University)
Han Sung-Hwan (Department of Chemistry, College of Natural Science, Hanyang University)
Koo Bon-Sung (Microbial Function Team, National Institute of Agricultural Biotechnology, RDA)
Yoon Moon-Young (Department of Chemistry, College of Natural Science, Hanyang University)
Publication Information
Korean Journal of Microbiology / v.41, no.4, 2005 , pp. 275-280 More about this Journal
Abstract
Anthrax is an infectious disease caused by the gram-positive bacterium, Bacillus anthracis. Anthrax toxin is a tripartite toxin comprising of protective antigen (PA), lethal factor (LF) and edema factor (EF). PA is the receptor-binding component, which facilitates the entry of LF or EF onto the cytosol. LF is a zinc-dependent metalloprotease, which is a critical virulence factor in cytotoxicity of infected animals. Therefore, it is of interest to develop its potent inhibitors for the neutralization of anthrax toxin. The first step to identify the inhibitors is the development of a rapid, sensitive, and simple assay method with a high-throughput ability. Much efforts have been concentrated on the preparation of powerful assays and on the screening of inhibitors using these system. In the present study, we have tried to construct anthrax lethal factor in yeast expression system to prepare cell-based high-throughput assay system. Here, we have shown the results covering the construction of a new vector system, subcloning of LF gene, and the expression of target gene. Our results are first trial to express LF gene in eukaryote and provide the basic steps in design of cell-based assay system.
Keywords
Bacillus anthracis; lethal factor; proteolytic activity; yeast expression system;
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