猪链球菌2型SSU05-1311蛋白的表达纯化和多克隆抗体的制备  

Gene expression,protein purification of SSU05-1311 from Streptococcus suis type 2 and preparation of the polyclonal antibody against the purified protein

作　　者：杨宝玲[1] 陈福广[2] 谢芳[2] 刘思国[2] 沈国顺[1] 张跃灵[2] 

机构地区：[1]沈阳农业大学畜牧兽医学院,辽宁沈阳110866 [2]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/动物细菌病研究室,黑龙江哈尔滨150001

出　　处：《中国兽医科学》2016年第3期310-314,共5页Chinese Veterinary Science

基　　金：国家自然科学基金青年科学基金项目(31500112)

摘　　要：以猪链球菌2型菌株05ZYH 33的基因组作为模板,PCR扩增SSU 05-1311(简称ssFbp)基因,克隆到表达载体pET22b中,构建重组质粒pET22b-ssFbp。该重组质粒转化到大肠杆菌BL21(DE3)中,用IPTG诱导表达SsFbp蛋白,并通过亲和层析纯化。纯化的SsFbp蛋白免疫BALB/c雌鼠,制备多克隆抗体。结果显示,Ss Fbp蛋白以可溶形式表达,分子质量大小为112ku。每升培养物获得SsFbp重组蛋白20 mg,纯度大于95%。免疫筛选后获得了ss Fbp蛋白的多克隆抗体。Western-blot结果显示,多克隆抗体能特异识别05ZYH33细胞壁蛋白组分中的SsFbp蛋白。本研究为进一步研究SsFbp蛋白的功能及其在致病性中的作用奠定了基础。The ss Fbp gene was amplified by PCR using the genomic DNA of Streptococcus suis type 205ZYH33 as template,and cloned into p ET22 b,yielding p ET22b-ss Fbp,which was transformed into E.coli BL21（DE3）. The Ss Fbp protein was expressed by inducing with IPTG and purified by affinity chromatography. Polyclonal antibody against Ss Fbp was was prepared by immunizing BALB/c mice.The results showed that Ss Fbp protein,with a molecular mass of 112 ku,was expressed as a soluble form. About 20 mg of Ss Fbp with purity above 95% were obtained from 1 L of culture. Polyclonal antibody against Ss Fbp was obtained. Western-blot analysis revealed that the polyclonal antibody specially recognized the natural Ss Fbp protein in the cell-wall protein fraction of 05ZYH33.The purified Ss Fbp and its antibody laid the foundation for further functional study on Ss Fbp protein.

关 键 词：猪链球菌2型 SsFbp蛋白 表达和纯化 多克隆抗体 

分 类 号：S852.612[农业科学—基础兽医学]