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原核表达幽门螺杆菌(Helicobacter pylori,H.pylori)酸适应感应蛋白CrdS,探讨其酸适应的调控机制。提取H.pylori26695标准株全基因组DNA作为模版,PCR扩增编码CrdS蛋白的基因hp1364,构建重组克隆质粒pUCm-T-hp1364和原核表达质粒pQE30-hp1364,经PCR、双酶切和测序鉴定正确后,转化大肠埃希菌XL1blue,IPTG诱导表达CrdS蛋白。结果表明:通过Western blot鉴定和SDS-PAGE分析该重组蛋白的相对分子质量约为46kDa,主要以包涵体形式存在。原核表达并成功获得的H.pylori CrdS蛋白,能够为探讨其酸适应的调控机制和新的抗H.pylori感染的途径提供实验材料。
Prokaryotic expression of Helicobacter pylori (H.pylori) acid-responsive protein CrdS, to explore its acid-adaptation regulation mechanism. The complete genome DNA of H.pylori26695 was extracted as a template to amplify hp1364 gene encoding CrdS protein by PCR. The recombinant plasmid pUCm-T-hp1364 and prokaryotic expression plasmid pQE30-hp1364 were constructed and identified by PCR, double enzyme digestion and sequencing , Transformed into Escherichia coli XL1blue, induced expression of CrdS protein by IPTG. The results showed that the molecular weight of the recombinant protein was about 46 kDa by Western blot and SDS-PAGE analysis, which was mainly in the form of inclusion bodies. The prokaryotic expression and successfully obtained H.pylori CrdS protein can provide experimental materials for exploring the mechanism of its acid adaptation and new anti-H.pylori infection pathways.