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IPTG诱导浓度、时间及温度对重组促性腺激素释放激素基因表达的影响
Effect of induce concentration, time and temperature of IPTG on the expression of the GST-GnRH/TRS gene
【摘要】 以人工合成的促性腺激素释放激素(gonadotropinreleasinghormone,GnRH)/转运肽(TRS)基因与表达载体pGEX-6p-1相连的重组子(pGE-G)作为研究对象,转化大肠杆菌BL21(ED3)plyS。利用IPTG作为诱导剂,分析比较不同IPTG诱导条件对于表达产物的影响,以确定最佳IPTG诱导条件。结果表明:融合蛋白大部分以包涵体的形式存在,诱导温度为30℃;IPTG浓度为0.2mmol/L时,可溶性融合蛋白GST-GnRH/TRS表达量最大。光密度扫描对表达产物进行初步定量,表明表达产物约占菌体总蛋白的33.3%。
【Abstract】 The object is recombinant GnRH2’S gene and transporter gene with fusion expression vector pGEX-6p-1, transformed into the BL21(ED3) plysS prokaryotic expression system, induced and expressed by IPTG, analyzed the effect of different concentration 、time and temperature of IPTG on the expression of the GST-GnRH/TRS gene, confirmed the best induced condition of IPTG. SDS-PAGE analysis showed that GST-GnRH/TRS expression quantity was the most when temperature of IPTG was 30 ℃ and concentration of IPTG was 0.2 mmol/L. The amount of the goal protein was evaluated by densitometric scanning. It indicated that the product of the GST-GnRH/TRS gene was 33% of total bacterial protein of BL21(ED3).
【Key words】 Gonadotropin releasing hormone(GnRH); induced concentration; exogenous protein; expression;
- 【文献出处】 黑龙江畜牧兽医 ,Heilongjiang Animal Science and Veterinary Medicine , 编辑部邮箱 ,2006年08期
- 【分类号】Q786
- 【被引频次】57
- 【下载频次】1751