【摘要】：目的分析表皮葡萄球菌SrrAB生物信息,表達及純化SrrA蛋白,為SrrA功能研究奠定基礎。方法以表皮葡萄球菌SE1457基因組為模板,PCR擴增srrA基因,構建重組表達質粒pET28a-srrA,轉入大腸桿菌BL21,利用純化的重組蛋白SrrA免疫小鼠,制備SrrA多克隆抗體,并檢測SrrA在表皮葡萄球菌不同生長時期的表達水平。序列比對利用Clustalw2軟件,蛋白結構域分析用DNAMAN軟件。結果表皮葡萄球菌SE1457SrrAB單獨組成一個操作子,SrrA位于胞漿中,與金黃色葡萄球菌和枯草桿菌類似物的同源性分別為95%和65%,SrrB為跨膜蛋白,與上述細菌類似物的同源性分別為71%和33%;表皮葡萄球菌SrrA于對數(shù)生長中期表達量最高。結論成功表達并純化了表皮葡萄球菌SrrA蛋白,為SrrAB下游調控機制研究奠定基礎。
[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.
【作者單位】： 大理大學基礎醫(yī)學院病原生物學綜合實驗室;復旦大學上海醫(yī)學院分子病毒學教育部/衛(wèi)生部重點實驗室;
【基金】：云南省科技廳應用基礎研究計劃項目(No.2014FB156) 大理大學高層次引進人才項目(No.KYBS201305) 云南省教學質量與教學改革工程資助項目-“白麗名師工作室”(云教高[2010]105號)~~
【分類號】：R378.1
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本文編號：2446188