| 摘要: |
| 目的：构建表达MafA(v-maf musculoaponeurotic fibrosarcoma oncogene homologue A)的逆转录病毒表达载体，并建立稳定表达MafA的肝原始细胞系(liver epithelial progenitor cells,LEPCs)。方法：通过PCR方法克隆MafA基因全长，将其构建到pBMN-Z-IRES-Neo逆转录病毒载体中获得pBMN-MafA-Neo载体；将该载体导入Phoenix包装细胞系,收集病毒上清并感染LEPCs,筛选稳定表达MafA的LEPCs；RT-PCR方法检测MafA表达对LEPCs分子表型的影响。结果：成功构建pBMN-MafA-Neo载体，并获得稳定表达MafA基因的肝原始细胞系(LEPCs-MafA)。LEPCs-MafA细胞GK和GLUT2基因表达高于LEPCs。结论：成功获得稳定表达MafA的肝原始细胞系，为研究MafA诱导肝干细胞向胰腺细胞转分化奠定了基础。 |
| 关键词: MafA 逆转录病毒载体 肝原始细胞系 |
| DOI：10.3724/SP.J.1008.2008.00465 |
| 投稿时间：2007-09-11修订日期：2008-01-21 |
| 基金项目:国家自然科学基金(30600326)，宁夏自然科学基金(NZ0775)，宁夏医学院特殊人才启动基金. |
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| Construction of MafA recombinant retroviral vector and its stable expression in liver epithelial progenitor cells |
| JIN Cai-xia1,2,LI Wen-lin1,XU Fang2*,HU Yi-ping1* |
| (1.Department of Cell Biology,College of Basic Medical Sciences,Second Military Medical University,Shanghai 200433,China;2.Department of Medical Genetics and Cell Biology,Ningxia Medical College,Yinchuan 750004) |
| Abstract: |
| Objective：To construct a recombinant retroviral vector harboring MafA and to establish a liver epithelial progenitor cell (LEPC) line for stable expression of MafA .Methods: MafA was amplified by PCR and suncloned into pBMN-Z-IRES-Neo vector to obtain pBMH-MafA-Neo vector.After introducing the pBMN-MafA-Neo into Phoenix package cells,the cell culture supernatant was used to infect LEPCs.LEPCs stably expressing MafA gene were screened out.RT-PCR was used to detect the influence of MafA on the phenotype of LEPCs.Results: We successfully constructed pBMH-MafA-Neo vector and obtained LEPCs which stably expressed MafA.Expression of GK and GLUT2 genes in LEPCs-MafA cells was higher than that in the LEPCs.Conclusion: We have successfully obtained LEPCs which can stably express MafA,laying a basis for studying the differentiation of LEPCs into pancreas cells. |
| Key words: MafA retroviral vector liver progenitor cells |
