粉尘螨Hsp60基因的测序、克隆和原核表达

Dermatophagoides farinae Hsp60--DNA sequencing,cloning and prokaryotic expression

目的探索和发现粉尘螨新的变应原组分。方法以粉尘螨热休克蛋白60(Heat shock protein 60,Hsp60)为研究对象,用简并引物PCR方法扩增出其cDNA片段,并对该片段进行了序列测定和分析。同时,对该基因片段进行了克隆、原核表达。结果与结论成功确定粉尘螨Hsp60基因片段cDNA序列;构建了pET-28a(+)-Hsp60重组质粒,并在E.coli BL21中得到高效表达。

We identified new Dermatophagoides farinae allergens by DNA sequencing,cloning and prokaryotic expression of Heat shock protein 60（Hsp60）fromDermatophagoides farinae.With degenerate primer polymerase chain reaction（PCR）to amplify the Dermatophagoides farinae cDNA,the cDNA fragments of the Hsp60 were abundantly obtained.The cDNA sequence and amino acid sequence encoded by it were detected and analysed.Meanwhile,the Hsp60 gene fragment was cloned into the pet-28 avector and transformed into E.coli for prokaryotic expression of the Hsp60.In this study,the cDNA sequence of Dermatophagoides farinae Hsp60 successfully was determined first,and the Hsp60 expressed in E.coli BL21.Results of this research laid a foundation for further analysis and study of Dermatophagoides farinae Hsp60.