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 p BV2 2 0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 p BV2 2 0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 2 0 % .结论 EPO模拟肽基因
AIM To clone and to express 4 repeats of EPO mimetic peptide gene. METHODS Whole EPO mimetic peptide gene was synthesized and inserted into pBS cloning vector. Multiple repeats of EPO mimetic peptide gene was gradually obtained with a new construction method. Then the correct gene repeats was transfered to Eco RI and Bam HI sites of pBV220 expression vector. RESULTS The sequencing and restriction analysis showed that 4 repeats of EPO mimetic peptide gene was correctly connected and was cloned into pBV220 vector. After the recombinant bacteria was induced at 42℃ for 4 h, a new anticipated protein band with relative molecule mass of 12 ku was found on SDS PAGE gel. The protein band amounted to 20% of total bacteria protein. CONCLUSION The correct 4 repeats of EPO mimetic peptide gene has successfully been constructed and expressed in E.coli .
出处
《第四军医大学学报》
北大核心
2001年第4期319-321,共3页
Journal of the Fourth Military Medical University
