人DcR3表达载体的构建及验证  

Over-expression vector construction of human DcR3 gene and its validation

作　　者：潘留兰[1] 贾胜男[1] 马静婷[1] 太京华[1] 金珍婧[1] 

机构地区：[1]吉林大学第二医院肝胆胰内科,长春130041

出　　处：《中国免疫学杂志》2016年第10期1491-1495,共5页Chinese Journal of Immunology

基　　金：吉林省科学技术委员会面上基金课题(201115083)

摘　　要：目的:构建人DcR3表达载体并验证其在体外细胞表达情况。方法:从人血液组织中扩增出DcR3中长度915 bp的CDS区,将其克隆到带有红色荧光的真核表达载体p EF1a-IRES-DsRed-Express2上,用Fu Gene HD转染法转染到肝星状细胞(LX-2)中,用RT-PCR以及Western blot检测其mRNA表达量和蛋白表达量。结果:通过RT-PCR和Western blot检测显示转染DcR3表达载体质粒的肝星状细胞中DcR3的转录和翻译水平均显著性提高。结论:成功构建出人DcR3表达载体并在LX-2细胞中高效表达。Objective： To construct the human DcR3 expression vector and verify its expression in vitro. Methods： 915 bp human DcR3 gene CDS was amplified from porcine lung tissues,and was cloned into eukaryotic expression vector p EF1a-IRES-DsRed-Express2 which show red fluorescence. And then p EF1a-IRES-DsRed-Express2-DcR3 was transfected into LX-2 cells by Fu Gene HD. Expression of mRNA and protein lever of Human DcR3 were detected by RT-PCR and Western blot. Results： The levels of DcR3 gene transcription and translation in the hepatic stellate cells were significantly increased after transfection with p EF1a-IRES-DsRed-Express2-DcR3 by RT-PCR and Western blot analysis. Conclusion： DcR3 expression vector was successfully constructed and highly expressed in LX-2 cells.

关 键 词：诱骗受体3 肝星状细胞 真核表达 克隆 

分 类 号：R392[医药卫生—免疫学]