【摘要】：目的分析表皮葡萄球菌SrrAB生物信息,表達(dá)及純化SrrA蛋白,為SrrA功能研究奠定基礎(chǔ)。方法以表皮葡萄球菌SE1457基因組為模板,PCR擴(kuò)增srrA基因,構(gòu)建重組表達(dá)質(zhì)粒pET28a-srrA,轉(zhuǎn)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(cè)SrrA在表皮葡萄球菌不同生長(zhǎng)時(shí)期的表達(dá)水平。序列比對(duì)利用Clustalw2軟件,蛋白結(jié)構(gòu)域分析用DNAMAN軟件。結(jié)果表皮葡萄球菌SE1457SrrAB單獨(dú)組成一個(gè)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(xì)菌類似物的同源性分別為71%和33%;表皮葡萄球菌SrrA于對(duì)數(shù)生長(zhǎng)中期表達(dá)量最高。結(jié)論成功表達(dá)并純化了表皮葡萄球菌SrrA蛋白,為SrrAB下游調(diào)控機(jī)制研究奠定基礎(chǔ)。
[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(xué)基礎(chǔ)醫(yī)學(xué)院病原生物學(xué)綜合實(shí)驗(yàn)室;復(fù)旦大學(xué)上海醫(yī)學(xué)院分子病毒學(xué)教育部/衛(wèi)生部重點(diǎn)實(shí)驗(yàn)室;
【基金】：云南省科技廳應(yīng)用基礎(chǔ)研究計(jì)劃項(xiàng)目(No.2014FB156) 大理大學(xué)高層次引進(jìn)人才項(xiàng)目(No.KYBS201305) 云南省教學(xué)質(zhì)量與教學(xué)改革工程資助項(xiàng)目-“白麗名師工作室”(云教高[2010]105號(hào))~~
【分類號(hào)】：R378.1
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本文編號(hào)：2446188