表皮葡萄球菌SrrAB生物信息分析及SrrA蛋白的原核表达
发布时间：2019-03-23 20:11
【摘要】：目的分析表皮葡萄球菌SrrAB生物信息,表达及纯化SrrA蛋白,为SrrA功能研究奠定基础。方法以表皮葡萄球菌SE1457基因组为模板,PCR扩增srrA基因,构建重组表达质粒pET28a-srrA,转入大肠杆菌BL21,利用纯化的重组蛋白SrrA免疫小鼠,制备SrrA多克隆抗体,并检测SrrA在表皮葡萄球菌不同生长时期的表达水平。序列比对利用Clustalw2软件,蛋白结构域分析用DNAMAN软件。结果表皮葡萄球菌SE1457SrrAB单独组成一个操作子,SrrA位于胞浆中,与金黄色葡萄球菌和枯草杆菌类似物的同源性分别为95%和65%,SrrB为跨膜蛋白,与上述细菌类似物的同源性分别为71%和33%;表皮葡萄球菌SrrA于对数生长中期表达量最高。结论成功表达并纯化了表皮葡萄球菌SrrA蛋白,为SrrAB下游调控机制研究奠定基础。
[Abstract]:Objective to analyze the biological information of Staphylococcus epidermidis SrrAB, express and purify SrrA protein, and lay a foundation for the study of SrrA function. Methods using the SE1457 genome of Staphylococcus epidermidis as template, the srrA gene was amplified by PCR, and the recombinant expression plasmid pET28a-srrA, was constructed. The recombinant expression plasmid pET28a-srrA, was transformed into E. coli BL21, to immunize mice with purified recombinant protein SrrA, and the polyclonal antibody to SrrA was prepared. The expression level of SrrA in different growth stages of Staphylococcus epidermidis was detected. Sequence alignment was performed by Clustalw2 software and protein domain analysis by DNAMAN software. Results Staphylococcus epidermidis SE1457SrrAB was composed of a single operator, and SrrA was located in cytoplasm. The homology with Staphylococcus aureus and Bacillus subtilis analogues was 95% and 65%, respectively. SRRB was a transmembrane protein. The homology with the above-mentioned bacterial analogues was 71% and 33%, respectively. The expression of Staphylococcus epidermidis SrrA was the highest in the middle of logarithmic growth. Conclusion the SrrA protein of Staphylococcus epidermidis was successfully expressed and purified, which laid a foundation for the study of downstream regulation mechanism of SrrAB.
【作者单位】： 大理大学基础医学院病原生物学综合实验室;复旦大学上海医学院分子病毒学教育部/卫生部重点实验室;
【基金】：云南省科技厅应用基础研究计划项目(No.2014FB156) 大理大学高层次引进人才项目(No.KYBS201305) 云南省教学质量与教学改革工程资助项目-“白丽名师工作室”(云教高[2010]105号)~~
【分类号】：R378.1
，
本文编号：2446188
[Abstract]:Objective to analyze the biological information of Staphylococcus epidermidis SrrAB, express and purify SrrA protein, and lay a foundation for the study of SrrA function. Methods using the SE1457 genome of Staphylococcus epidermidis as template, the srrA gene was amplified by PCR, and the recombinant expression plasmid pET28a-srrA, was constructed. The recombinant expression plasmid pET28a-srrA, was transformed into E. coli BL21, to immunize mice with purified recombinant protein SrrA, and the polyclonal antibody to SrrA was prepared. The expression level of SrrA in different growth stages of Staphylococcus epidermidis was detected. Sequence alignment was performed by Clustalw2 software and protein domain analysis by DNAMAN software. Results Staphylococcus epidermidis SE1457SrrAB was composed of a single operator, and SrrA was located in cytoplasm. The homology with Staphylococcus aureus and Bacillus subtilis analogues was 95% and 65%, respectively. SRRB was a transmembrane protein. The homology with the above-mentioned bacterial analogues was 71% and 33%, respectively. The expression of Staphylococcus epidermidis SrrA was the highest in the middle of logarithmic growth. Conclusion the SrrA protein of Staphylococcus epidermidis was successfully expressed and purified, which laid a foundation for the study of downstream regulation mechanism of SrrAB.
【作者单位】： 大理大学基础医学院病原生物学综合实验室;复旦大学上海医学院分子病毒学教育部/卫生部重点实验室;
【基金】：云南省科技厅应用基础研究计划项目(No.2014FB156) 大理大学高层次引进人才项目(No.KYBS201305) 云南省教学质量与教学改革工程资助项目-“白丽名师工作室”(云教高[2010]105号)~~
【分类号】：R378.1
，
本文编号：2446188
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