中国人兽共患病学报2017，Vol.33Issue(3)：250-255,6.DOI:10.3969/j.issn.1002-2694.2017.03.011
2型猪链球菌FtsZ重组蛋白的制备及其酶活性测定
Purification and enzyme activity assay of filamentous temperature-sensitive protein Z in Streptococcus suis serotype 2
摘要
Abstract
We conducted purification of filamentous temperature-sensitive protein Z of Streptococcus suis serotype 2 (S.suis 2) and measured its GTPase activity.The ftsz gene in the genome of the Chinese 05ZYH33 strain of S.suis 2 was successfully amplified using PCR,and then the ftsz gene was cloned into prokaryotic expression plasmid pET28a,and the recombinant plasmid pET28a-ftsz was transformed into E.coli BL21.After induction by IPTG,the isolated FtsZ protein was analyzed with SDS-PAGE.Then the recombinant protein was purified by Ni2+-NTA affinity chromatography.The rabbit serum was harvested after immunization with recombinant FtsZ protein,and was analyzed by indirect ELISA and Western blotting.The GTPase activity of FtsZ was measured with the malachite green method.Results showed that successfully constructed recombinant plasmid pET28a-ftsz and the recombinant protein with high purity was obtained;Western blot result indicated that FtsZ could react with the His-tag antibody and the rabbit serum;the polyclonal antibody titer of the rabbit serum reached 1 ∶ 13 107 200;FtsZ have GTPase activity.We successfully prepared S.suis 2 recombinant protein FtsZ having GTPase activity and high titer antiserum would be useful for the further study of S.suis 2 cell division mechanism.关键词
2型猪链球菌/丝状温度敏感蛋白/纯化Key words
Streptococcus suis serotype 2/filamentous temperature-sensitive protein Z/purification分类
医药卫生引用本文复制引用
范伟伟,倪华,孙卫平,张剑,李超龙,吴倩倩,王长军,潘秀珍..2型猪链球菌FtsZ重组蛋白的制备及其酶活性测定[J].中国人兽共患病学报,2017,33(3):250-255,6.基金项目
国家自然科学基金(No.81571965,No.81471920)、江苏省自然科学基金(No.BK20151091)、江苏省333工程科研资助项目(No.BRA2014363)联合资助 （No.81571965,No.81471920）
Funded by the National Natural Science Foundation of China (Nos.81571965 & 81471920),the Natural Science Foundation of Jiangsu Province (No.BK20151091),and the 333 Engineering Science Foundation of Jiangsu Province (No.BRA2014363) （Nos.81571965 & 81471920）
