改变部分TIR对EPO模拟肽8串联体表达的影响  

Modified DNA sequence of translation initiation region helps improve expression of 8 repeats of EPO mimetic peptide

作　　者：韩苇[1] 颜真[1] 王俊楼[1] 赵永同[1] 石继红[1] 张英起[1] 

机构地区：[1]第四军医大学生物技术中心,陕西西安710033

出　　处：《第四军医大学学报》2001年第3期224-226,共3页Journal of the Fourth Military Medical University

摘　　要：目的　通过改变部分翻译起始区序列观察对 EPO模拟肽基因 8串联体表达的影响 .方法　设计和人工合成了部分翻译起始区域 (translation initiation region,TIR) DNA序列 ,将该序列插入 p BSEPOM8(含 EPO模拟肽基因 8串联体序列 )的 Eco RI和 X ba I位点 ,经 DNA测序确定正确重组质粒 ,再将 EPO模拟肽基因 8串联体 (含改建的翻译起始区域 )克隆到 p BV2 2 0载体的 Eco RI和 Bam HI位点 ,诱导表达 .结果　 DNA测序证明 ,人工合成的 DNA序列已正确插入EPO模拟肽基因 8串联体的翻译起始区域 .该串联体基因插入 p BV2 2 0载体后 ,经 42℃诱导 ,在细菌裂解上清中出现一条相对分子质量为 2 2× 10 3(Mr)的新蛋白带 ,与 EPO模拟肽基因 8串联体的理论计算分子量相符合 .经薄层扫描证明该蛋白带约占细菌蛋白总量的 15 % .结论　通过改变部分翻译起始区域起动了 EPO模拟肽基因 8串联体的翻译 ,使原来不表达的 EPO模拟肽基因AIM To observe effect of a synthesized DNA sequence of translation initiation region (TIR) on expression of 8 repeats of EOP mimetic peptide gene. METHODS A DNA sequence of translation initiation region was designed, synthesized and inserted into Eco RI and Xba I sites of pBSEPOM 8 which contained 8 repeats of EPO mimetic peptide gene. The correct recombinant plasmid DNA was named for pBSEPOM 8B. Then 8 repeats of EPO mimetic peptide gene which contained synthesized TIR sequence was cloned into Eco RI and Bam HI sites of pBV220 expression vector. RESULTS DNA sequencing analysis showed that the sequence of synthesized TIR was correctly inserted into Eco RI and Xba I sites of pBVEPOM 8B. After recumbent bacterium contained pBVEPOM 8B was induced at 42℃ for 4 h, a new protein band was found on SDS PAGE gel. The relative molecule mass of the new protein band was about 22 000 ( M r), consistent with 8 repeats of EPO mimetic peptide. The protein band amounted to 15% of total bacteria protein. CONCLUSION Synthesized DNA sequence of TIR initiates translation of 8 repeats of EPO mimetic peptide gene, and so helps improve the previously non expression of the latter.

关 键 词：翻译起始区域 SD序列 EOP模拟肽 基因串联体 

分 类 号：Q78[生物学—分子生物学]