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

[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.