HO-1基因真核表达载体pcDNA3.1-hHO-1的构建及表达  

Construction and expression of eukaryotic expression vector of hHO-1 gene

作　　者：王媛[1] 金国平[1] 马惠敏[1] 单杰[1] 

机构地区：[1]郑州大学基础医学院生物化学与分子生物学教研室,河南郑州450001

出　　处：《河南医学研究》2009年第1期19-22,共4页Henan Medical Research

基　　金：郑州市科委资助项目(064S6D33218-32)

摘　　要：目的:构建人HO-1真核表达载体pcDNA3.1-hHO-1并检测其表达。方法:利用分子生物学技术,将hHO-1基因与质粒pcDNA3.1连接,构成重组质粒并用RT-PCR及Western blotting方法检测重组载体在Eca-9706细胞中的表达情况,最终确定重组载体是否构建成功。结果:用HindⅢ和BamHⅠ双酶切,RT-PCR检测,Western blotting检测证明hHO-1成功克隆入表达载体pcDNA3.1中。结论:本部分实验成功构建了能够在细胞内大量表达HO-1基因产物的真核表达载体pcDNA3.1-hHO-1,为进一步实验奠定基础。Objective ： To construct eukaryotic expression vector pcDNA3.1-hHO-1 of homo sapiens HO-1 gene and detect its expression. Methods： The hHO-1 gene is inserted into eukaryotic expression vector pcDNA3.1. After it show that the insertion is successful by digesting with Hind Ⅲ ＋ BamH Ⅰ , we transfect the Eca-9706 cell line with this reorganized vector. We detect the expression of HO-1 in cells transfected with reorganized vector using RT-PCR and western blotting methods to decide whether or not the reorganized eukaryotic expression vector is successfully constructed. Results ： Digesting with HindⅢ ＋ BamH Ⅰ , it proves that the HO-1 gene has been cloned into eukaryotic expression vector pcDNA3.1. RT-PCR and western blotting results suggest that the reorganized vector has worked well in the cells and expressed great quantity of HO-1 product. Conclusion ： We have construct a eukaryotic expression vector pcDNA3.1-hHO-1 of homo sapiens HO- 1 gene successfully.

关 键 词：血红素加氧酶-1 Eca-9706细胞 pcDNA3.1质粒 

分 类 号：R318[医药卫生—生物医学工程]