嗜肺军团菌热休克蛋白60基因的克隆及序列分析

Cloning and Sequencing of Heat Shock Protein 60(HSP60) Gene of Legionella Pneumophila

目的获取嗜肺军团菌热休克蛋白60(HSP60)基因,并将它克隆到质粒pUC18中进行核苷酸序列分析。方法应用聚合酶链式反应(PCR)技术从嗜肺军团菌扩增得到HSP60基因,将其定向插入载体pUC18并转化大肠杆菌JM109,用限制性酶切及PCR对重组质粒进行鉴定。用双脱氧链末端终止法进行DNA序列测定,并与GenBank中报道的HSP60基因进行比较。结果成功克隆了约1647bp的HSP60基因片段,测序结果表明,所克隆的HSP60基因序列与GenBank公布的一致性为98%。结论获得了序列正确的HSP60基因,为其重组表达及相关研究奠定了基础。

Objective To obtain the heat shock protein 60(HSP60) gene of Legionella pneumophila and clone it into plasmid pUC18 for nucleotide sequencing. Methods The HSP60 gene of Legionella pneumophila was amplified by PCR, then inserted into pUC18 vector and transformed into Escherichia coli(E.coli)strain JM109. The recombinant plasmids were identified by restriction analysis and PCR. HSP60 gene was sequenced by the dideoxy chain termination method, then analyzed and compared with that reported in GenBank. Results The HSP60 gene of 1647 bp in length was successfully cloned. Comparison of the sequence of HSP60 gene with that reported in GenBank showed that the identities were 98%. Conclusion A confirmed HSP60 gene has been obtained in this paper.The study laid a good foundation of the recombination,expression and research of the gene.