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Construction of adenoviral expression vector carrying FLIP gene and its expression in rat pulmonary vascular endothelial cells
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 18
- DOI:
- No doi available
- Key Word:
- FLIP;腺病毒;肺血管内皮细胞;ALI/ARDS
Abstract： 目的:构建FLIP(FLICE-inhibitory protein)腺病毒表达载体并观察该载体在大鼠肺血管内皮细胞(PVEC)中的表达情况,为研究FLIP蛋白的抗凋亡作用打下基础. 方法:利用含有大鼠FLIP基因的原始载体,通过Admax腺病毒包装系统,构建表达大鼠FLIP基因的重组腺病毒Ad-FLIP;原代培养并鉴定大鼠PVEC.利用成功构建的重组腺病毒Ad-FLIP感染大鼠PVEC.通过逆转录酶一多聚酶链反应(reverse tran-seriptase-polymerase chain reaction,RT-PeR)和Western Blot分别检测经重组腺病毒Ad-FLIP感染后大鼠PVEC及对照组细胞中FLIP基凶mRNA和蛋白表达水平. 结果:成功构建含有大鼠FLIP基因的重组腺病毒Ad-FIJP;经Ⅷ因子间接免疫荧光鉴定原代培养大鼠PVEC纯度达90%以上;大鼠PVEC经重组腺病毒Ad-FLIP感染后24 h即检测到FLIP mRNA和蛋白表达水平明显增高. 结论:重组腺病毒Ad-FLIP可有效提高大鼠PVEC中FLIP基因的表达水平.
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