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载脂蛋白E3的克隆及原核表达

The Cloning of Apolipoprotein E3 Gene and Its Expression in E.coli
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摘要 The apoE cDNA was amplified from four months old fetus liver tissue mRNA by RT PCR. It was inserted into the TOPO ○R vector, whose sequence is the same as that reported. The right apoE cDNA was inserted into pQE30 vector to construct pQE30 apoE recombinant expressed plasmid, which was expressed as a histidine tag fusion protein in E.coli strain BL21(DE3) with higher level. The purified protein was gained by Ni NTA column. SDS PAGE showed that 34 kD protein was expressed. Western blot analysis showed a positive band at 34 kD. The apoE cDNA was amplified from four months old fetus liver tissue mRNA by RT PCR. It was inserted into the TOPO ○R vector, whose sequence is the same as that reported. The right apoE cDNA was inserted into pQE30 vector to construct pQE30 apoE recombinant expressed plasmid, which was expressed as a histidine tag fusion protein in E.coli strain BL21(DE3) with higher level. The purified protein was gained by Ni NTA column. SDS PAGE showed that 34 kD protein was expressed. Western blot analysis showed a positive band at 34 kD.
出处 《中国生物化学与分子生物学报》 CAS CSCD 北大核心 2003年第1期133-135,共3页 Chinese Journal of Biochemistry and Molecular Biology
关键词 载脂蛋白E3 克隆 原核表达 human apoE 3 cDNA, molecular cloning, gene expression
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