正反义人Tankyrase基因真核表达载体的构建及鉴定  

Construction and identification of sense and antisense human Tankyrase eukaryotic expression vector

作　　者：杨孟华[1] 房殿春[1] 杨仕明[1] 罗元辉[1] 刘为纹[1] 

机构地区：[1]第三军医大学西南医院全军消化中心,重庆400038

出　　处：《第四军医大学学报》2003年第13期1224-1226,共3页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金 (30 1 70 4 2 5)

摘　　要：目的 :构建人Tankyrase正义和反义真核表达载体 .方法 :用EcoRⅠ从TT2 0质粒上切下约 3.4 4kb的TankyrasecDNA片段 ,然后连入pcDNA3.1/Zeo的EcoRⅠ酶切位点上 ,经BamHⅠ酶切鉴定出正义和反义表达载体 .结果 :经BamHⅠ酶切后 ,正义质粒形成 5 .0 +3.4kb两条带 ,而反义重组质粒为 8.2 +0 .2kb两条带 ,与理论计算值完全一致 .结论 :成功构建了人Tankyrase的正。AIM: To construct a eukaryotic expressing vector of sense and antisense human Tankyrase gene. METHODS : The human Tankyrase cDNA fragment contained in the TT20 vector was cloned into the eukaryotic expressing vector pcDNA3.1 in cis direction or trans direction using DNA recombinant technology. The recombinant vector was further identified by digestion of Bam H Ⅰ. RESULTS: After digested by Bam HⅠ, two fragments which lengthened 5.0 and 3.4 kb were formed in sense eukaryotic expressing vector (pcDNA TNKS), while two fragments which lengthened 8.2 kb and 0.2 kb were formed in antisense vector (pcDNA aTNKS). Electrophoretic results were completely coincident with theoritical calculation. CONCLUSION: Human Tankyrase sense and antisense eukaryotic expressing vectors are successfully constructed.

关 键 词：TANKYRASE 端粒酶 反义基因 真核表达载体 基因表达 鉴定 正义基因 

分 类 号：Q78[生物学—分子生物学]