Pdx-1真核表达载体构建及其在大鼠骨髓间充质干细胞中的表达

Construction eukaryotic expression vector of Pdx-1 and expression in rat mesenchymal stem cells

目的 构建含有胰腺-十二指肠同源盒-1(Pdx-1)的真核表达载体,并研究其在大鼠骨髓间充质干细胞(MSCs)中的表达情况.方法 克隆Pdx-1基因,构建含Pdx-1的真核表达载体pEGFP-Pdx-1及pcDNA3.1-Pdx-1,转染MSCs,倒置荧光显微镜及逆转录聚合酶链反应(RT-PCR)检测MSCs中Pdx-1的表达.结果 测序结果 表明克隆的Pdx-1基因序列正确,构建的含Pdx-1的真核表达载体能有效的转染MSCs,并检测到该基因mRNA的表达.结论 完成Pdx-1基因克隆,成功构建含有Pdx-1基因真核表达载体,并在转化株中检测到该基因的表达,为糖尿病的细胞替代及基因治疗提供基础.

Objective To construct eukaryotic expression vector containing the pancreatic-duodenum homeobox-1(Pdx-1)and to study the expression in rat bone marrow-derived mesenchymal stem cells (MSCs). Methods Cloned Pdx-1 gene and constructed containing Pdx-1 eukaryotic expression vector pEGFP-Pdx-1 and pcDNA3.1-Pdx-1, transfected MSCs, detected the Pdx-1 expression in MSCs by invert fluorescence microscope and RT-PCR.Results Sequencing results showed that the cloned Pdx-1 gene sequence was correct, the eukaryotic expression vector containing Pdx-1 could effectively transfected MSCs and detected the gene expression at mRNA level.Conclusions The completion of Pdx-1 gene is cloned successfully and constructed containing Pdx-1 gene eukaryotic expression vector, detected the expression of the gene in the stable transfected MSCs,making preparations for the cell replacement and gene therapy of diabetes.