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Effect of SeaR gene on virginiamycins production in Streptomyces virginiae

희소방선균 SeaR 유전자가 Streptomyces virginiae의 virginiamycins 생산에 미치는 영향

  • Ryu, Jae-Ki (Department of Biomedical Laboratory Science, Gimcheon University) ;
  • Kim, Hyun-Kyung (Department of Biomedical Laboratory Science, Gimcheon University) ;
  • Kim, Byung-Won (Department of Biomedical Laboratory Science, Gimcheon University) ;
  • Kim, Dong-Chan (Department of Biomedical Laboratory Science, Gimcheon University) ;
  • Lee, Hyeong-Seon (Department of Biomedical Laboratory Science, Jungwon University)
  • 류재기 (김천대학교 임상병리학과) ;
  • 김현경 (김천대학교 임상병리학과) ;
  • 김병원 (김천대학교 임상병리학과) ;
  • 김동찬 (김천대학교 임상병리학과) ;
  • 이형선 (중원대학교 임상병리학과)
  • Received : 2015.07.30
  • Accepted : 2015.09.01
  • Published : 2015.09.30

Abstract

In order to study the effect of the receptor protein (SeaR), which is isolated from Saccharopolyspora erythraea, we introduced the SeaR gene to Streptomyces virginiae as host strains. An effective transformation procedure for S. virginiae was established based on transconjugation by Escherichia coli ET12567/pUZ8002 with a ${\varphi}C31$-derived integration vector, pSET152, which contained int, oriT, attP, and $ermEp^{\ast}$ (erythromycin promotor). Therefore, the pEV615 was introduced into S. virginiae by conjugation and integrated at the attB locus in the chromosome of the recipients by the ${\varphi}C31$ integrase (int) function. Transformants of S. virginiae containing the SeaR gene were confirmed by PCR and transcriptional expression of the SeaR gene in the transformants was analyzed by RT-PCR, respectively. And, we examined the production time of virginiamycins in the culture media of both the transformants and the wild type. The production time of virginiamycins in the wild type and transformants was the same. When 100 ng/ml of synthetic $VB-C_6$ was added to the state of 6 or 8 hour cultivation of wild type and transformants, respectively, the virginiamycins production was induced, meaning that the virginiamycins production in the wild type was detected 2 h early than transformants. From these results, SeaR expression was also affected to virginiamycins production in transformants derived from S. virginiae. In this study, we showed that the SeaR protein worked as a repressor in transformants.

Acknowledgement

Supported by : 김천대학교

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