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KMID : 0364820150510030256
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Korean Journal of Microbiology
2015 Volume.51 No. 3 p.256 ~ p.262
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Effect of SeaRgene on virginiamycins production in Streptomyces virginiae
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Ryu Jae-Ki
Kim Hyun-Kyung
Kim Byung-Weon
Kim Dong-Chan
Lee Hyeong-Seon
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Abstract
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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 φC31-derived integration vector, pSET152, which contained int, oriT, attP, and ermEp* (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 φC31 integrase (int) function. Transformants of S. virginiae containing the SeaRgene were confirmed by PCR and transcriptional expression of the SeaRgene 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/㎖ of synthetic VB-C6 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 SeaRprotein worked as a repressor in transformants.
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KEYWORD
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Saccharopolyspora erythraea, SeaRgene, Streptomyces virginiae, transformant, virginiamycin
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