Abstract:To identify and demonstrated the properties of Rgg transcription regulator in highly virulent strains of Streptococcus suis serotype 2, the rgg gene from the genomic DNA in the virulent strain 05ZYH33 was amplified by PCR and subcloned into plasmid pMD18-T and pQE30 with double digestion of BamHⅠand Hind Ⅲ. Subsequently, the prokaryotic expression plasmid pQE30-rgg was transformed to E. coli M15 after identification by restriction endonuclease digestion and DNA sequencing. Upon induction with IPTG, E. coli M15 containing the recombinant plasmid could express a distinct band with a molecular weight of 30 kDa, which was similar to the predicted band of Rgg protein as demonstrated by SDS-PAGE and Western blot. It was demonstrated that the recombinant protein expression could be the highest soluble yield after induction for overnight at 15 ℃. The recombinant Rgg protein was purified by Ni2+ NTA affinity chromatography. Native-PAGE results showed that the purified protein forms homodimers in vitro, consistent with data for other members of the Rgg family. Secondary structure analysis displayed Rgg protein contained 15 α-helices and 2 β-turn. These results would be useful in the further studies on the function of Rgg transcription regulator.
郑峰, 刘鹏, 蔡炳冈, 朱进, 潘秀珍, 王长军. 2型猪链球菌Rgg调控因子的序列结构分析及原核表达[J]. 中国人兽共患病学报, 2016, 32(3): 229-233.
ZHENG Feng, LIU Peng, CAI Bing-gang, ZHU Jin, PAN Xiu-zhen, WANG Chang-jun. Sequence and structure analysis of Rgg transcription regulator in Streptococcus suis serotype 2 and prokaryotic expression. Chinese Journal of Zoonoses, 2016, 32(3): 229-233.
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