Discrimination of Bacillus subtilis from other Bacillus Species Using Specific Oligonucleotide Primers for the Pyruvate Carboxylase and Shikimate Dehydrogenase Genes

Lee Gawon; Heo Sojeong; Kim Tao; Na Hong-Eun; Park Junghyun; Lee Eungyo; 이종훈; 정도원

doi:10.4014/jmb.2205.05014

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자료유형: 학술저널

저자정보: Lee Gawon (Department of Food and Nutrition Dongduk Women’s University Seoul 02748 Republic of Korea) Heo Sojeong (Department of Food and Nutrition Dongduk Women’s University Seoul 02748 Republic of Korea) Kim Tao (Department of Food and Nutrition Dongduk Women’s University Seoul 02748 Republic of Korea) Na Hong-Eun (Department of Food and Nutrition Dongduk Women’s University Seoul 02748 Republic of Korea) Park Junghyun (Department of Food and Nutrition Dongduk Women’s University Seoul 02748 Republic of Korea) Lee Eungyo (Department of Food and Nutrition Dongduk Women’s University Seoul 02748 Republic of Korea) 이종훈 (경기대학교) 정도원 (동덕여자대학교)

저널정보: 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제32권 제8호

발행연도: 2022.8

수록면: 1,011 - 1,016 (6page)

DOI: 10.4014/jmb.2205.05014

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Bacillus subtilis is a useful bacterium in the food industry with applications as a starter strain for fermented food and as a probiotic. However, it is difficult to discriminate B. subtilis from other Bacillus species because of high phenotypic and genetic similarity. In this study, we employed five previously constructed multilocus sequence typing (MLST) methods for the discrimination of B. subtilis from other Bacillus species and all five MLST assays clearly distinguished B. subtilis. Additionally, the 17 housekeeping genes used in the five MLST assays also clearly distinguished B. subtilis. The pyruvate carboxylase (pyrA) and shikimate dehydrogenase (aroE) genes were selected for the discrimination of B. subtilis because of their high number of polymorphic sites and the fact that they displayed the lowest homology among the 17 housekeeping genes. Specific primer sets for the pyrA and aroE genes were designed and PCR products were specifically amplified from B. subtilis, demonstrating the high specificity of the two housekeeping genes for B. subtilis. This species-specific PCR method provides a quick, simple, powerful, and reliable alternative to conventional methods in the detection and identification of B. subtilis.

#Bacillus subtilis #16S rRNA gene #multilocus sequence typing #pyrA #aroE

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