本文選題：幽門螺桿菌 + HspA��； 參考：《重慶醫(yī)科大學(xué)》2010年碩士論文

【摘要】： 目的: 構(gòu)建含幽門螺桿菌(Hp)熱休克蛋白A編碼基因的重組載體,并電轉(zhuǎn)入乳酸乳球菌MG1363,表達(dá)目的蛋白并分析其免疫反應(yīng)性,為Hp基因工程口服疫苗的研究和開發(fā)奠定基礎(chǔ)。 方法: 以Hp NCTC11637菌株基因組DNA為模板,PCR擴(kuò)增hspA基因,并克隆至乳酸乳球菌表達(dá)載體pMG36e中。將重組質(zhì)粒轉(zhuǎn)化E.coli DH5ɑ,經(jīng)鑒定的陽性重組質(zhì)粒命名為pMG36e/hspA。以電穿孔法將pMG36e/hspA轉(zhuǎn)化乳酸乳球菌MG1363并用Western blotting檢測HspA蛋白的表達(dá)。 結(jié)果: 成功構(gòu)建重組質(zhì)粒pMG36e/hspA。將pMG36e/hspA電轉(zhuǎn)化MG1363后,收集菌體蛋白進(jìn)行Western blotting分析,在HspA的相對分子質(zhì)量(Mr≈13kDa)處出現(xiàn)特異性條帶,顯示表達(dá)的重組蛋白具有良好的免疫反應(yīng)性。 結(jié)論: 首次成功構(gòu)建了表達(dá)Hp HspA的重組乳酸乳球菌MG1363,為進(jìn)一步口服疫苗的相關(guān)研究奠定了基礎(chǔ)。
[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，

本文編號：1929930