摘要
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.
出处
《安徽农学通报》
2017年第8期12-15,共4页
Anhui Agricultural Science Bulletin
基金
吉林省科技发展计划资助项目(20130101105JC
20140204018YY)
吉林农业大学博士科研启动基金项目(2015015)