结核分枝杆菌RD1区Rv3873基因真核表达载体的构建及鉴定  

CONSTRUCTION AND IDENTIFICATION OF MYCOBACTERIUM TUBERCULOSIS Rv3873 FROM RD1 EUKARYOTIC EXPRESSION VECTOR

作　　者：曾林子[1] 陈建平[2] 刘成君[1] 李红霞[2] 姚卫[3] 杨志荣[1] 

机构地区：[1]四川大学生命科学院生物资源与生态环境教育部重点实验室,成都610065 [2]华西医学中心基础医学与法医学院寄生虫教研室 [3]四川绵阳市疾病预防控制中心

出　　处：《现代预防医学》2007年第11期2001-2003,共3页Modern Preventive Medicine

基　　金：国家自然科学基金资助项目(30300302);四川省学术和技术带头人培养基金(4200316)

摘　　要：[目的]构建结核分枝杆菌RD1区Rv3873基因的真核表达载体pcDNA3.1-Rv3873,并对其进行鉴定。[方法]利用PCR技术从结核杆菌H37Rv中扩增Rv3873基因,并将其定向克隆至原核载体pGEX-4T-1中,经测序鉴定后,将重组质粒pGEX-4T-1-Rv3873中的目的基因亚克隆入真核表达载体pcDNA3.1(+),构建重组子pcDNA3.1-Rv3873,通过限制性内切酶切酶及PCR进行鉴定。[结果]克隆的Rv3873基因序列与GenBank公布的一致性为100%,限制性内切酶酶切及PCR分析表明Rv3873已成功插入pcDNA3.1(+)中。[结论]成功构建了Rv3873基因真核表达载体,为进一步研究新型结核杆菌DNA疫苗奠定了基础。[Objective] To construct and identify the eukaryotic expression vector of Mycobacterium tuberculosis Rv3873. [ Methods ] Rv3873 was amplified by PCR and directly cloned into expression plasmid pGEX-4T-1 for sequencing. The target gene in the recombinant plasmid （pGEX-4T-1-Rv3873） was sub-cloned into the eukaryotic expression vector pcDNA3.1 （＋） and the recombinant pcDNA3.1-Rv3873. This recombinant plasmid was identified by restriction analysis and PCR. [Results] Comparison of the sequence of Rv3873 gene with that reported in GenBank showed that the identities were 100%.And it was comfirmed that Rv3873 gene was successfully inserted into pcDNA3.1 （＋） . [Conclusion] We successfully construct the eukaryotic expression vector pcDNA3.1-Rv3873 and it lays a good foundation for the research of new DNA vaccine of Mycobacterium tubereulosis.

关 键 词：结核分枝杆菌 RD1 Rv3873 真核表达载体 

分 类 号：R378.911[医药卫生—病原生物学]