Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Isolation and Characterization of Some Promoter Sequences from Leuconostoc mesenteroides SY2 Isolated from Kimchi

  • Park, Ji Yeong (Industrial Technology Research Group, World Institute of Kimchi (an Annex of Korea Food Research Institute)) ;
  • Jeong, Seon-Ju (College of Pharmacy, Catholic University of Daegu) ;
  • Kim, Jeong A (Division of Applied Life Science (BK21 Plus), Graduate School, Gyeongsang National University) ;
  • Kim, Jeong Hwan (Division of Applied Life Science (BK21 Plus), Graduate School, Gyeongsang National University)
  • Received : 2017.03.16
  • Accepted : 2017.06.26
  • Published : 2017.09.28
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Abstract

Some promoters were isolated and characterized from the genome of Leuconostoc mesenteroides SY2, an isolate from kimchi, a Korean traditional fermented vegetable. Chromosomal DNA of L. mesenteroides SY2 was digested with Sau3AI and ligated with BamHI-cut pBV5030, a promoter screening vector containing a promoterless cat-86. Among E. coli transformants (TFs) resistant against Cm (chloramphenicol), 17 were able to grow in the presence of $1,000{\mu}g/ml$ Cm and their inserts were sequenced. Transcription start sites were examined for three putative promoters (P04C, P25C, and P33C) by primer extension. Four putative promoters were inserted upstream of a promoterless ${\alpha}$-amylase reporter gene in $pJY15{\alpha}$. ${\alpha}$-Amylase activities of E. coli TFs containing $pJY15{\alpha}$ (control, no promoter), $pJY03{\alpha}$ ($pJY15{\alpha}$ with P03C), $pJY04{\alpha}$ (with P04C), $pJY25{\alpha}$ (with P25C), and $pJY33{\alpha}$ (with P33C) were 66.9, 78.7, 122.1, 70.8, and 99.3 U, respectively. Cells harboring $pJY04{\alpha}$ showed 1.8 times higher activity than the control. Some promoters characterized in this study might be useful for construction of food-grade expression vectors for Leuconostoc sp. and related lactic acid bacteria.

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References

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