本文選題：布魯氏菌 + WbdA基因��； 參考：《江蘇農(nóng)業(yè)學(xué)報(bào)》2017年02期

【摘要】：研究分析羊種布魯氏菌043新疆流行株中編碼脂多糖的WbdA基因和WbkE基因的原核表達(dá)以及蛋白的生物信息學(xué)信息。以043菌株為模板,構(gòu)建重組質(zhì)粒pET-30a-WbdA和pET-30a-WbkE,轉(zhuǎn)化至E.coli BL21(DE3)菌株,IPTG誘導(dǎo)表達(dá),然后通過Western Blot分析檢測(cè)目的蛋白的表達(dá),最后運(yùn)用DNAMAN和TMHMM Server v.2.0軟件等對(duì)WbdA基因和WbkE基因編碼的氨基酸序列進(jìn)行分析。結(jié)果顯示,成功構(gòu)建了WbdA基因和WbkE基因的原核表達(dá)載體pET-30a-WbdA和pET-30a-WbkE,并且在大腸桿菌中成功表達(dá)。生物信息學(xué)分析結(jié)果顯示,043菌株的WbdA蛋白和WbkE蛋白與標(biāo)準(zhǔn)菌株16M的WbdA蛋白和WbkE蛋白的同源性高。WbdA蛋白有1個(gè)跨膜結(jié)構(gòu)區(qū),沒有信號(hào)肽,20個(gè)抗原決定簇,二級(jí)結(jié)構(gòu)主要由α-螺旋構(gòu)成,利用Phyre2在線軟件成功構(gòu)建了該蛋白的3D模型。WbkE蛋白沒有跨膜結(jié)構(gòu)區(qū),沒有信號(hào)肽,有17個(gè)抗原決定簇,二級(jí)結(jié)構(gòu)主要由α-螺旋構(gòu)成,并利用Phyre2在線軟件成功構(gòu)建了該蛋白的3D模型。
[Abstract]:The prokaryotic expression of WbdA gene and WbkE gene encoding lipopolysaccharide and the bioinformatics information of protein of Brucella brucella 043 in Xinjiang were studied. Using strain 043 as template, the recombinant plasmids pET-30a-WbdA and pET-30a-WbkE were constructed and transformed into E.coli BL21DE3 strain. The expression of the target protein was detected by Western Blot analysis. Finally, the amino acid sequences of WbdA gene and WbkE gene were analyzed by DNAMAN and TMHMM Server v.2.0 software. The results showed that the prokaryotic expression vectors pET-30a-WbdA and pET-30a-WbkE of WbdA gene and WbkE gene were successfully constructed and successfully expressed in Escherichia coli. Bioinformatics analysis showed that the homology of WbdA protein and WbkE protein was high with that of standard strain 16M WbdA protein and WbkE protein. WbdA protein had one transmembrane structure region, no signal peptide and 20 antigenic determinants. The secondary structure is mainly composed of 偽 -helix. The 3D model of the protein. WbkE protein has no transmembrane structure region, no signal peptide, and 17 antigenic determinants. The secondary structure is mainly composed of 偽 -helix. The 3D model of the protein was successfully constructed by using Phyre2 online software.
【作者單位】： 新疆石河子大學(xué)生命科學(xué)學(xué)院;新疆石河子大學(xué)動(dòng)物科技學(xué)院;
【基金】：國家“973”計(jì)劃項(xiàng)目(2010CB530200)
【分類號(hào)】：S852.61

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