摘要：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. Abstract： 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.

关键词：真核表达载体构建 大鼠 骨髓间充质干细胞 mesenchymal stem 

单位：郑州大学第一附属医院内分泌及代谢疾病科 450052 河南省人民医院老年临床医学部内分泌科 郑州大学生物工程系