人幽门螺杆菌HspA亚单位的基因克隆及序列分析VIP免费

人幽门螺杆菌 HspA 亚单位的基因克隆及序列分析【摘要】 目的： 获取幽门螺杆菌（Hp）热休克蛋白 A（ HspA）亚单位基因并进行核苷酸序列分析和同源性比较。方法： 获取幽门螺杆菌的染色体 DNA，PCR 技术扩增获取 HspA 基因，将其定向插入pGEX-4T-1 原核表达载体中进行核苷酸序列分析。结果： 经酶切鉴定及基因序列证实 Hp HspA 基因克隆成功，DNA 序列与 Genbank 公布的序列比较有 13 个碱基存在差异，同源性为 96%；由碱基序列推定编码的氨基酸残基序列没有发生变异，同源性为 100%。结论： 成功地将 Hp HspA 基因克隆到了pGEX 4T-1 原核表达载体，为 HspA 的表达和研究奠定了实验基础。 【关键词】 螺杆菌，幽门； HspA 亚单位； 克隆，生物；基因顺序 ［Abstract］ Objective: To get heat shock protein A （HspA）subunit gene of Helicobacter pylori, analyze its DNA sequence, and compare its homology. Methods: Chromosome DNA of H.pylori was extracted and the HspA gene was amplified with Polymerase Chain Reaction （PCR）. The obtained DNA fragment was cloned into prokaryotic expression vector pGEX-4T-1 and sequenced. Results: HspA gene was cloned successfully which was demonstrated by enzyme digesting and gene sequencing methods. Comparing with DNA sequences reported in Genbank, nucleotide sequence of the obtained DNA fragment was 96% identical with that of HspA gene with a variation of 13 base pairs. The homology in putative amino acids was 100%. Conclusions: HspA gene has been Cloned into prokaryotic expression vector pGEX 4T-1 successfully, which makes an experimental foundation for its expression and associated research. ［Key words］ Helicobacter pylori；HspA subunit；cloning, organism; gene order 幽门螺杆菌（Helicobacter pylori,Hp）是上消化道的重要致病菌之一，其感染与 B 型胃炎和消化性溃疡的发生密切相关，并被世界卫生组织列为胃癌发生的相关致病菌［1］。在 Hp 的免疫治疗研究方面，纯化的尿素酶（Ure）和热休克蛋白（Heat shock protein，Hsp）及...