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

发布时间：2018-05-01 02:19

本文选题：猪链球菌型 + MCP　；参考：《中国人兽共患病学报》2013年03期

【摘要】：目的原核表达猪链球菌2型甲基受体趋化蛋白编码基因05SSU0273。方法通过PCR方法检测该基因在不同血清型猪链球菌中的分布情况,然后利用获得的猪链球菌2型05SSU0273基因片段,构建重组表达载体pET32a::05SSU0273。将质粒转入E.coli BL21中,经IPTG诱导,SDS-PAGE和Western blot实验证实目的基因的表达,然后以His亲和层析柱纯化重组蛋白。结果在S.suis 2 35个血清型标准株中,有20株分离株扩增出目的条带。重组质粒可在宿主菌中高效表达,经镍柱亲和层析得到相对分子质量为43kD的重组蛋白。结论本实验成功获得S.suis 2MCP蛋白,为研究其在猪链球菌2型致病过程中发挥的作用奠定了基础。
[Abstract]:Objective to express the encoding gene 05SSU0273 of methyl-receptor chemoattractant protein of Streptococcus suis type 2 in prokaryotic expression. Methods the distribution of the gene in different serotype Streptococcus suis was detected by PCR method. Then the recombinant expression vector pET32a: 05SSU0273 was constructed by using the obtained 05SSU0273 gene fragment of Streptococcus suis type 2. The plasmid was transferred into E.coli BL21. The expression of the target gene was confirmed by IPTG induced SDS-PAGE and Western blot. The recombinant protein was purified by His affinity chromatography. Results out of the 235 serotype standard strains of S.suis, 20 strains were isolated and the target bands were amplified. The recombinant plasmid was highly expressed in the host strain and the recombinant protein with relative molecular weight of 43kD was obtained by nickel column affinity chromatography. Conclusion S.suis 2MCP protein was successfully obtained in this study, which laid a foundation for the study of its role in the pathogenicity of Streptococcus suis type 2.
【作者单位】：南京师范大学生命科学学院;南京军区军事医学研究所;
【基金】：国家自然科学基金(No.81071317;No.81172794;No.30972638;No.31170124) 江苏省自然科学基金项目(No.BK2010113;No.BK2011098;No.BK2010025;No.BK2010114;No.BK2011097)资助~~
【分类号】：S852.611;R378.1

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