关键词: 模拟肽;基因表达;大肠杆菌

    摘 要:目的 克隆和表达EPO模拟肽基因4串联体. 方法 设计了特殊的基因串联方案，在人工合成EPO模拟肽全基因的基础上，将EPO模拟肽基因由单体逐步地连接成4串联体.继而将获得的正确EPO模拟肽4串联体插入pBV220表达载体诱导表达. 结果 构建的EPO模拟肽基因4串联体经酶切和DNA测序分析，结果表明该基因串联体的序列与设计的序列完全相同.EPO模拟肽4串联体插入pBV220载体后，重组菌经42℃诱导4h，SDS-PAGE分析结果显示，该串联体得到较高水平的表达.凝胶扫描结果表明，其表达量约占菌体蛋白总量的20%. 结论 EPO模拟肽基因4串联体的构建与表达均获得了成功.

    Construction and expression of4repeats of EPO mimetic peptide gene

    HAN Wei，YAN Zhen，WANG Jun-Lou，ZHAO Yong-Tong，SHI Ji-Hong，ZHANG Ying-Qi

    Biotechnology Center，Fourth Military Medical U-niversity，Xi'an710033，China

    Keywords:mimetic peptide;gene expression;Escherichia coli

    Abstract:AIM To clone and to express4repeats of EPO mimetic peptide gene.METHODS Whole EPO mimetic pep-tide gene was synthesized and inserted into pBS cloning vec-tor.Multiple repeats of EPO mimetic peptide gene was grad-ually obtained with a new construction method.Then the correct gene repeats was transfered to EcoRI and BamHI sites of pBV220expression vector.RESULTS The sequencing and restriction analysis showed that4repeats of EPO mimetic peptide gene was correctly connected and was cloned into pBV220vector.After the recombinant bacteria was induced at42℃for4h ......