羊种布鲁氏菌043新疆流行株WbdA基因和WbkE基因的原核表达及生物信息学分析

发布时间：2018-06-04 00:45

  本文选题：布鲁氏菌 + WbdA基因　； 参考：《江苏农业学报》2017年02期

【摘要】：研究分析羊种布鲁氏菌043新疆流行株中编码脂多糖的WbdA基因和WbkE基因的原核表达以及蛋白的生物信息学信息。以043菌株为模板,构建重组质粒pET-30a-WbdA和pET-30a-WbkE,转化至E.coli BL21(DE3)菌株,IPTG诱导表达,然后通过Western Blot分析检测目的蛋白的表达,最后运用DNAMAN和TMHMM Server v.2.0软件等对WbdA基因和WbkE基因编码的氨基酸序列进行分析。结果显示,成功构建了WbdA基因和WbkE基因的原核表达载体pET-30a-WbdA和pET-30a-WbkE,并且在大肠杆菌中成功表达。生物信息学分析结果显示,043菌株的WbdA蛋白和WbkE蛋白与标准菌株16M的WbdA蛋白和WbkE蛋白的同源性高。WbdA蛋白有1个跨膜结构区,没有信号肽,20个抗原决定簇,二级结构主要由α-螺旋构成,利用Phyre2在线软件成功构建了该蛋白的3D模型。WbkE蛋白没有跨膜结构区,没有信号肽,有17个抗原决定簇,二级结构主要由α-螺旋构成,并利用Phyre2在线软件成功构建了该蛋白的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 伪 -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 伪 -helix. The 3D model of the protein was successfully constructed by using Phyre2 online software.
【作者单位】： 新疆石河子大学生命科学学院;新疆石河子大学动物科技学院;
【基金】：国家“973”计划项目(2010CB530200)
【分类号】：S852.61

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