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%。
The object is recombinant GnRH2S 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 ).
出处
《黑龙江畜牧兽医》
CAS
北大核心
2006年第8期17-19,共3页
Heilongjiang Animal Science And veterinary Medicine
关键词
促性腺激素释放激素
诱导浓度
外源蛋白
表达
Gonadotropin releasing hormone(GnRH)
induced concentration
exogenous protein
expression