幽门螺杆菌热休克蛋白A在乳酸乳球菌中的表达及免疫反应性鉴定

发布时间：2018-05-24 17:29

  本文选题：幽门螺杆菌 + HspA　； 参考：《重庆医科大学》2010年硕士论文

【摘要】： 目的: 构建含幽门螺杆菌(Hp)热休克蛋白A编码基因的重组载体,并电转入乳酸乳球菌MG1363,表达目的蛋白并分析其免疫反应性,为Hp基因工程口服疫苗的研究和开发奠定基础。 方法: 以Hp NCTC11637菌株基因组DNA为模板,PCR扩增hspA基因,并克隆至乳酸乳球菌表达载体pMG36e中。将重组质粒转化E.coli DH5ɑ,经鉴定的阳性重组质粒命名为pMG36e/hspA。以电穿孔法将pMG36e/hspA转化乳酸乳球菌MG1363并用Western blotting检测HspA蛋白的表达。 结果: 成功构建重组质粒pMG36e/hspA。将pMG36e/hspA电转化MG1363后,收集菌体蛋白进行Western blotting分析,在HspA的相对分子质量(Mr≈13kDa)处出现特异性条带,显示表达的重组蛋白具有良好的免疫反应性。 结论: 首次成功构建了表达Hp HspA的重组乳酸乳球菌MG1363,为进一步口服疫苗的相关研究奠定了基础。
[Abstract]:Objective: A recombinant vector containing heat shock protein A (HPA) encoding gene of Helicobacter pylori was constructed and transferred into Lactococcus lactis MG1363. The recombinant vector expressed the target protein and analyzed its immunoreactivity, which laid a foundation for the research and development of oral HP gene engineering vaccine. Methods: The hspA gene was amplified by genomic DNA of HP NCTC11637 strain and cloned into the expression vector pMG36e of Lactococcus lactis. The recombinant plasmid was transformed into E.coli DH5. The identified positive recombinant plasmid was named pMG36e / hspA. PMG36e/hspA was transformed into Lactococcus lactis MG1363 by electroporation and the expression of HspA protein was detected by Western blotting. Results: The recombinant plasmid pMG36e / hspA was successfully constructed. After pMG36e/hspA was electrotransformed into MG1363, specific bands were found at the relative molecular weight of HspA, Mr 鈮，

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