pHBAd-MCMV-GFP-NCAM1真核表达质粒的构建和鉴定  

Construction and identification of eukaryotic expression plasmid p HBAd-MCMV-GFP-NCAM1

作　　者：李永明[1] 陈立强[1] 张东东[1] 赵勇[1] 李晓涛[2] 李艳君[1] 

机构地区：[1]佳木斯大学基础医学院,黑龙江佳木斯154007 [2]佳木斯大学第一附属医院,黑龙江佳木斯154007

出　　处：《解剖学研究》2015年第1期22-25,共4页Anatomy Research

基　　金：黑龙江省自然科学基金(C201212)

摘　　要：目的将目的基因NCAM1与质粒载体p HBAd-MCMV-GFP进行连接,得到重组质粒p HBAd-MCMV-GFPNCAM1。方法对目的基因和质粒载体分别用内切酶Not I和Nsil进行双酶切,产物回收后进行连接、转化,得到重组质粒p HBAd-MCMV-GFP-NCAM1。转化后的菌液进行PCR阳性克隆鉴定,并对阳性样本进行测序。结果首先对NCAM1进行扩增,从实验结果看符合预期效果,在双酶切、连接和转化实验后,对构建完成的重组质粒进行鉴定,PCR阳性克隆鉴定结果显示重组质粒构建成功,NCAM1已连接进入重组质粒中,测序的结果进一步印证阳性鉴定结果。结论带有目的基因NCAM1的重组质粒构建成功。Objective Connected the NCAM1 gene and p HBAd-MCMV-GFP to the recombinant plasmid p HBAd-MCMVGFP-NCAM1.Methods The target genes NCAM1 and the vector p HBAd-MCMV-GFP respectively was digested by Not I and Nsil.The recovered product was connected and transformed to the recombinant plasmid p HBAd-MCMV-GFP-NCAM1.The recombinant plasmid was identified by PCR and sequencing.Results uccess amplify the gen NCAM1;The gen NCAM1 was connected into the recombinant plasmid by PCR and sequencing.Conclusions The recombinant plasmid p HBAd-MCMV-GFPNCAM1 was successfully constructed.

关 键 词：神经细胞黏附分子1 质粒载体 重组质粒 

分 类 号：R346[医药卫生—基础医学]