摘要
通过原核表达系统表达猪链球菌Sao-M和Ide_(Ssuis)基因,并分析其反应原性。根据GenBank数据库发表的序列,利用分子生物学软件设计了2对特异性引物,通过PCR扩增Sao-M和Ide_(Ssuis)基因,经测序分析后,分别克隆到pET28a(+)和pMAL-c2X,构建重组表达质粒pET28a:Sao-M和p MAL-c2X:Ide_(Ssuis),然后将鉴定为阳性的重组质粒分别转入宿主菌BL21和Rosetta2中用IPTG进行诱导表达,通过免疫印迹进行鉴定。实验表明,两种重组蛋白在原核系统中获得了高效的表达,并能与相应的阳性抗血清发生特异性反应。本研究为进一步探究Sao-M和Ide_(Ssuis)蛋白生物学功能及其在猪链球菌致病机制上的作用提供了依据。
Prokaryotic expression system was used to express Sao- M and Ide_（Ssuis）of Streptococcus suis（ S. suis）,and the reactionogenicity was analyzed. Two pairs of specific primers were designed according to sequences downloaded from Gen Bank to amplify those two genes by using polymerase chain reaction（ PCR） and the product of PCR was sequenced and subcloned into pET28a（ ＋） and p MAL- c2 X respectively,getting two recombinant plasmids named pET28a： Sao- M and p MAL- c2X： Ide_（Ssuis）. Positive recombinant plasmids were transformed into BL21 and Rosetta 2 and then the recombinant strains were induced by IPTG. Western blotting was used to detect the recombinant proteins. Results showed that the two recombinant proteins were expressed with high efficiency and solubility,and could react specifically with the positive antiserum of S. suis. This study provides a reference for the further research of the biological function of Sao- M and Ide_（Ssuis）and Streptococcus universal vaccines.
出处
《中国兽药杂志》
北大核心
2016年第6期1-4,共4页
Chinese Journal of Veterinary Drug
基金
山东省自然科学基金资助项目(ZR2013CQ004)
