pdx-1基因克隆及其在人肝癌细胞系SMMC-7721中的表达  

Cloning the Gene Pancreatic Duodenal Homeobox-1 and Its Expression in SMMC-7721 Hepatoma Cells

作　　者：陈元晓[1] 李文林[2] 何志颖[2] 巴月[3] 田明[1] 訾晓渊[2] 张彦[1] 胡以平[2] 

机构地区：[1]昆明医学院细胞生物学及遗传学教研室,云南昆明650031 [2]第二军医大学细胞生物学教研室,上海200433 [3]郑州大学公共卫生学院,河南郑州450052

出　　处：《癌变．畸变．突变》2006年第2期81-83,共3页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家自然科学基金资助(No.30270668;30200138)

摘　　要：背景与目的克隆小鼠胰十二指肠同源框-12(PEGFP-C1pdx-1)基因,并构建pdx-1的表达载体。材料与方法通过RT-PCR方法从小鼠胰腺总RNA中克隆pdx-1基因,并将该基因构建到pEGFP-C1和pcDNA3.0中获得pdx-1的表达载体。通过观察荧光和RT-PCR方法检测表达载体转染SMMC-7721后pdx-1基因在细胞中的表达。结果克隆了883bp的pdx-1全长cDNA,成功构建了pEGFP-C1pdx-1和pcDNA3pdx-1两种表达载体。pEGFP-C1pdx-1转染SMMC-7721后,可观察到绿色荧光仅局限于细胞核中;通过RT-PCR方法在转染后的细胞中都检测到了pdx-1基因的转录。结论pdx-1表达载体的构建为进一步研究pdx-1基因在胰腺发育和肝干细胞增殖与分化方面的作用奠定了基础。BACKGROUND ＆ AIM： To clone the mouse pdx-1 gene and to construct its expression vector. MATERIAL AND METHODS： Mouse pdx-1 gene was amplified from mouse pancreas total RNA and subcloned into the vectors pEGFP-C1 and peDNA3.0. The expression of pdx-1 was analyzed by RT-PCR and green fluorescence detection after transfection of the SMMC-7721 cell line. RESULTS： The expression vectors of pEGFP-C1 pdx-1 and peDNA3 pdx-1 were constructed. After transfecting the SMMC-7721 cells with pEGFP-C1 pdx-1, green fluorescence was detected only in the nucleoli. By RT-PCR analysis, pdx-1 mRNA was detected in both pEGFP-C1 pdx-1 and pcDNA3 pdx-1 transfected cells. CONCLUSION： The expression vectors of pdx-1 gene were useful to investigate the development of the pancreas and the proliferation and differentiation of hepatic stem cells.

关 键 词：胰十二指肠同源框-1(pdx-1) 基因克隆 基因表达 

分 类 号：Q782[生物学—分子生物学]