Purification and enzyme activity assay of filamentous temperature-sensitive protein Z in Streptococcus suis serotype 2
10.3969/j.issn.1002-2694.2017.03.011
- VernacularTitle:2型猪链球菌FtsZ重组蛋白的制备及其酶活性测定
- Author:
Weiwei FAN
;
Hua NI
;
Weiping SUN
;
Jian ZHANG
;
Chaolong LI
;
Qianqian WU
;
Changjun WANG
;
Xiuzhen PAN
- Keywords:
Streptococcus suis serotype 2;
filamentous temperature-sensitive protein Z;
purification
- From:
Chinese Journal of Zoonoses
2017;33(3):250-255
- CountryChina
- Language:Chinese
-
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.
