Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Discrimination of Bacillus subtilis from Other Bacillus Species Using Specific Oligonucleotide Primers for the Pyruvate Carboxylase and Shikimate Dehydrogenase Genes

  • Lee, Gawon (Department of Food and Nutrition, Dongduk Women's University) ;
  • Heo, Sojeong (Department of Food and Nutrition, Dongduk Women's University) ;
  • Kim, Tao (Department of Food and Nutrition, Dongduk Women's University) ;
  • Na, Hong-Eun (Department of Food and Nutrition, Dongduk Women's University) ;
  • Park, Junghyun (Department of Food and Nutrition, Dongduk Women's University) ;
  • Lee, Eungyo (Department of Food and Nutrition, Dongduk Women's University) ;
  • Lee, Jong-Hoon (Department of Food Science and Biotechnology, Kyonggi University) ;
  • Jeong, Do-Won (Department of Food and Nutrition, Dongduk Women's University)
  • Received : 2022.05.11
  • Accepted : 2022.07.05
  • Published : 2022.08.28
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Abstract

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.

Keywords

Acknowledgement

This work was supported by the National Research Foundation of Korea (NRF) [NRF-2019R1A2C1003639]. We thank Edanz (https://www.edanz.com/ac) for editing a draft of this manuscript.

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