中国人兽共患病学报2013，Vol.29Issue(3)：220-225,6.DOI:10.3969/cjz.j.issn.1002-2694.2013.03.003
猪链球菌2型甲基受体趋化蛋白编码基因05SSU0273的原核表达及纯化
Prokaryotic expression and purification of the recombinant methyl-accepting chemotaxis protein gene 05SSU0273 from Streptococcus suis serotype 2
摘要
Abstract
To study the biological function of the methyl-accepting chemotaxis protein (MCP) gene 05SSU0273 and its products in Streptococcus suis serotype 2 , a pair of specific primers was designed for the gene 05SSU0273 , and the target DNA fragment was successfully amplified using the genomic template of Chinese strains 05ZYH33 . Subsequently , target gene was inserted into pMD18-T vector, and then subcloned into prokaryotic expression vector pET32a, generating a recombinant expression plasmid pET32a:: 05SSU0273 . Direct DNA sequencing method was used to confirm its correctness . The recombinant plasmid was then transformed into E. coli BL21. After IPTG induction , this bacteria could produce the target MCP protein with 6 X his-tag which could be purified by affinity chromatography system . The results of SDS-PAGE and Western-blot showed that the fusion protein had a relative molecular weight of 43 kD . Moreover, MCP gene had been detected in 20 of 35 S. suis strains with different serotypes . This prevalence indicated that protein MCP might play a role in the survival of S. suis 2 in different hosts . Together, our work laid foundation for functional research on MCP in S. suis 2 .关键词
猪链球菌2型/MCP/原核表达Key words
Streptococcus suis type 2 / MCP/ prokaryotic expressionc分类
医药卫生引用本文复制引用
郭静静,杜骁杰,刘文静,郑峰,胡丹,王长军,潘秀珍..猪链球菌2型甲基受体趋化蛋白编码基因05SSU0273的原核表达及纯化[J].中国人兽共患病学报,2013,29(3):220-225,6.基金项目
国家自然科学基金(No.81071317 （）
No.81172794 （）
No.30972638 （）
No.31170124)和江苏省自然科学基金项目(No.BK2010113 （）
No.BK2011098 （）
No.BK2010025 （）
No.BK2010114 （）
No.BK2011097)资助 （）
