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海洋科学  2011 

中国对虾细胞色素P450基因CYP4原核表达条件优化

, PP. 49-55

Keywords: 中国对虾(Fenneropenaeus,chinensis),细胞色素P450,原核表达

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Abstract:

根据克隆得到的中国对虾(Fenneropenaeuschinensis)CYP4基因开放阅读框设计引物,构建了中国对虾CYP4基因原核表达载体p28a-CYP4,并对重组菌株Rosetta/28a-CYP4的原核表达条件进行了优化。结果表明:p28a-CYP4转化Rosetta后可实现CYP4基因的原核表达,SDS-PAGE分析显示其在56.0kDa处有显著诱导条带;通过对诱导温度、IPTG浓度、诱导时机(OD600)及诱导时间的优化表明,重组菌株Rosetta/p28a-CYP4的最佳诱导温度为37℃,最佳IPTG浓度为1.2mmol/L,最佳诱导时机及诱导时间分别为0.59和6h。

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