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表达Kallistatin的9型重组腺相关病毒的制备及其体外表达效率

DOI: 10.11830/ISSN.1000-5013.2013.01.0068

Keywords: 9型重组腺相关病毒, Kallistatin, 表达效率, 人脐静脉血管内皮细胞

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Abstract:

采用三质粒磷酸钙共转染生产系统,制备出可以表达内源性抗肿瘤血管生成因子(Kallistatin)的9型重组腺相关病毒(rAAV2/9)和2型重组腺相关病毒(rAAV2/2).rAAV经银染法鉴定其纯度;采用Western Blotting法比较rAAV2/9和rAAV2/2外壳蛋白的大小;qPCR方法检测rAAV的滴度.使用rAAV2/9-Kallistatin和rAAV2/2-Kallistatin分别感染人脐静脉血管内皮细胞(HUVEC)后,RT-PCR和ELISA法分别检测Kallistatin的mRNA和蛋白表达水平.结果表明:纯化的病毒纯度达到98%以上;rAAV9外壳蛋白比rAAV2的大.感染HUVEC后, rAAV2/9介导Kallistatin在mRNA和蛋白表达水平都要比rAAV2/2高.

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