结核分枝杆菌Rv3872与Rv3873基因的克隆及重组真核表达载体的构建

Cloning of Mycobacterium tuberculosis Rv3872 and Rv3873 Genes and Construction of Recombinant Expression Vector

目的:克隆结核分枝杆菌Rv3872与Rv3873基因,并构建Rv3872与Rv3873基因的重组真核表达载体,为研究这2个基因的功能奠定实验基础。方法:利用PCR技术克隆Rv3872与Rv3873基因序列,将其连接至pMD-18T载体,鉴定成功后将Rv3872与Rv3873序列插入真核表达载体pEGFP-C1,构建重组pEGFP-C1-Rv3872和pEGFP-C1-Rv3873真核表达载体,并进行质粒PCR、双酶切以及测序验证。结果:经测序比对后,Rv3872与Rv3873序列与Gene Bank中公布的基因序列一致,重组载体构建成功。结论:成功构建重组pEG-FP-C1-Rv3872和pEGFP-C1-Rv3873真核表达载体。

Objective：Cloning of Mycobacterium tuberculosis Rv3872 and Rv3873 gene and constructing recombinant eukaryotic expression vector of Rv3872 and Rv3873 gene.Methods： The whole gene of Rv3872 and Rv3873 sequence were cloned by PCR, and then the gene were inserted into the pMD-18T, then inserted it into the pEGFP-C1 after successful validation, constructing the recombinant eukaryotic expression vectors of pEGFP-C 1-Rv3872 and pEGFP-C1-Rv3873, and then the plasmid was validated by PCR and double enzyme digestion and sequencing.Resluts ： Rv3872 and Rv3873 were consistent with the gene sequences published in GeneBank after sequencing, and the recombinant vector was successfully constructed.Conclusion：The recombinant eukaryotic expression vectors pEGFPC1-Rv3872 and pEGFP-C1-Rv3873 were successfully constructed.