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Characterization of antimicrobial proteins produced by Bacillus sp. N32  

Lee, Mi-Hye (Microbial Genetics Division, National Institute of Agricultural Biotechnology (NIAB))
Park, In-Cheol (Microbial Genetics Division, National Institute of Agricultural Biotechnology (NIAB))
Yeo, Yun-Soo (Microbial Genetics Division, National Institute of Agricultural Biotechnology (NIAB))
Kim, Soo-Jin (Microbial Genetics Division, National Institute of Agricultural Biotechnology (NIAB))
Yoon, Sang-Hong (Microbial Genetics Division, National Institute of Agricultural Biotechnology (NIAB))
Lee, Suk-Chan (Department of Genetic Engineering, Sungkyunkwan University)
Chung, Tae-Young (Department of Genetic Engineering, Sungkyunkwan University)
Koo, Bon-Sung (Microbial Genetics Division, National Institute of Agricultural Biotechnology (NIAB))
Publication Information
The Korean Journal of Pesticide Science / v.10, no.1, 2006 , pp. 56-65 More about this Journal
Abstract
An antagonistic bacterial isolate, that inhibits the growth of plant pathogens, was selected and identified from 5,000 isolates screened from the rhizosphere of various crop plants. An isolate Bacillus sp. N32, tested against Colletotrichum gloeosporioides causing anthracnose disease in hot pepper, produced both a heat resistant antifungal protein and a heat sensitive antifungal protein. The heat resistant protein was partially purified by Ammonium sulfate fractionation and gel filtration chromatography. The bioautography showed that the proteins possessed high antifungal activity. The biosynthetic gene cluster responsible for the heat resistant antifungal protein was cloned from cosmid library using DNA probe obtained from PCR product with the primers targeting the conserved nucleotide sequence of the synthetic genes reported earlier, Most of the clones obtained showed higher homology to fengycin antibiotic synthetic gene family reported earlier. On the other hand, the heat sensitive protein was isolated from SDS-PAGE and electroblotting to determine the N-terminal amino acid sequences. The heat sensitive antifungal protein gene was cloned from the ${\lambda}-ZAP$ libraries using a DNA probe based on the N-terminal amino acid sequences of the heat sensitive protein. We are contemplating to clone and sequence the whole gene cluster encoding the heat sensitive protein for further analysis.
Keywords
antifungal peptide; anthracnose disease; biosynthetic gene; Colletotrichum gloeosporioides; cosmid library; fengycin; Rhizobacteria; ${\lambda}-ZAP$ library;
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