恙虫病东方体Karp株Sta22抗原基因原核表达载体的构建及鉴定

Construction and identification of prokaryotic expression vector including Sta22 gene of Orientia tsutsugamushi

目的:构建恙虫病东方体Karp株Sta22抗原基因原核表达载体,为表达该基因奠定基础。方法:以恙虫病东方体Karp株基因组为模板,通过PCR方法扩增Sta22抗原基因,并将其克隆到表达载体pEASY-E2上,通过PCR方法及核苷酸测序进行鉴定。结果:所构建的原核表达载体携带Sta22基因,基因序列与GenBank上所公布的信息完全一致。结论:成功构建了Sta22抗原基因的原核表达载体,为研制和开发新型的恙虫病疫苗及临床诊断试剂奠定理论基础及物质保障。

Objective：To construct the prokaryotic expression vector including Sta22 gene from Orientia tsutsugamushi,and provide foundation for further study of Sta22 gene expression.Methods： Orientia tsutsugamushi karp genome was used as template to amplify Sta22 gene by polymerase chain reaction,Sta22 gene was cloned into the prokaryotic expression vector pEASY-E2 and recombinant involved Sta22 gene was then identified.Results：The results proved that the construction of prokaryotic expression vector included Sta22 gene.DNA sequence analyzing revealed that the Sta22 gene sequences were totally consistent with information retrieved from the GenBank.Conclusions：The prokaryotic expression vector containing Sta22 gene is successfully constructed,and we provide the theoretical and material foundation for further study of vaccine and clinical diagnostic reagents of scrub typhus.