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冷诱导rna结合蛋白(cirp)过表达载体的构建及鉴定

DOI: 10.3724/SP.J.1145.2014.08010, PP. 381-384

Keywords: 冷诱导rna结合蛋白,过表达,质粒,冷应激

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Abstract:

为构建携带冷诱导rna结合蛋白(coldinduciblerna-bindingprotein,cirp)的过表达载体,首先从4℃条件下冷处理8h的sd大鼠睾丸组织中获得cirp基因的cdna序列,然后扩增cirp基因的编码区,将其克隆到真核表达载体plenti6/v5中,酶切鉴定并测序.将克隆成功的质粒转染hek293t细胞,用westernblot检测cirp的表达水平.成功扩增cirp编码区,并将其克隆至载体plenti6/v5中;当将cirp过表达载体转染hek293t细胞后,westernblot检测结果显示cirp蛋白表达水平明显增加(p<0.01).本研究成功构建了cirp过表达载体,为进一步研究cirp的作用机制提供了基础工具.

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