激活素受体相互作用蛋白ARIPzip真核表达载体的构建及表达  

Construction of eukaryotic vector of activin receptor interacting protein ARIPzip and expression in HEK293 cells

作　　者：裴颖[1] 刘飙[2] 鲍永利[3] 于春雷[3] 孙颖[3] 李玉新[3] 乌垠[3] 苏冠方[1] 

机构地区：[1]吉林大学第二医院眼科,吉林长春130041 [2]吉林大学中日联谊医院手外科,吉林长春130033 [3]东北师范大学药物基因和蛋白筛选国家工程实验室,吉林长春130024

出　　处：《中国实验诊断学》2009年第3期293-295,共3页Chinese Journal of Laboratory Diagnosis

摘　　要：目的构建激活素受体相互作用蛋白(activin receptor interacting protein zip,ARIPzip)真核表达载体,以便进一步探讨其生物学作用。方法从小鼠成纤维细胞3T3中提取RNA,并利用ARIPzip特异性引物通过RT-PCR方法扩增了ARIPzip cDNA,并克隆入pcDNA-Flag载体中,构建重组载体pcDNA-F-ARIPzip。将重组载体转染入HEK293细胞中,并通过免疫印迹方法检测ARIPzip的表达。结果酶切及测序结果显示,重组质粒构建成功;免疫印迹结果显示,瞬时转染重组质粒的HEK293细胞中可有效表达ARIPzip。结论本研究成功构建了激活素受体相互作用蛋白ARIPzip真核表达载体,并能够在真核细胞中进行表达,为进一步研究ARIPzip的生物学功能奠定了基础。Objective Construct the activin receptor interacting protein AR1Pzip recombinant eukaryofic expression vector, in order to further investigate the biological functuion of ARIP-Ap.Methods Extract total RNA from-mouse fibroblast cell line 3T3 and amplify ARIPzip eDNA by RT-PCR using specific primers. Then the cDNA was inserted into vector pcDNA-Flag to construct the recombinant vector pcDNA-F-ARIPzip,then the recombinant vector was tmusfected into HEK293 cells and the protein expression was detected by western blot. Results Enzyme restriction and sequencing data indicated that the recombinant vector was constructed exactly, western blot result shown that the ARIPzip recombinanteukaryotic expression vector can efficiently express ARlPzip in HEK293 cells. Conclusion The eukaryotic expression vector pcDNA-F-ARIPzip is conctracted and expresses in HEK293 cells successfully, which establish the foundation for future research on ARIPzip gene function.

关 键 词：激活素受体相互作用蛋白 ARIPzip 真核表达 HEK293细胞 

分 类 号：Q78[生物学—分子生物学]