摘要
目的 :对人IFNα_2b基因的 5′端进行改造 ,从翻译水平上调控IFNα_2b基因在大肠杆菌中的表达。方法 :采用PCR方法 ,人工合成寡核苷酸引物 ,对人IFNα_2b基因进行了改造 :去除3′端非编码区 ,并增加原核系统强终止密码子。将改造后的基因片段分别克隆入原核表达载体pBV32 1,在大肠杆菌中进行表达。对表达产物进行活性效价测定。结果 :3′端删除非编码区 ,表达水平比删除前提高 4倍。结论 :IFNα_2b基因的 3′端非编码区对其在原核系统的表达具有抑制作用 ,删除该区可提高表达水平。
To regulate the expression level of HuIFNα_2b gene in E. coli by modifying its 5′ terminal. Method:PCR method was used to synthesize oligonucleotide primers,and HuIFNα_2b gene was modified as follows:3′ noncoding region was removed,and a potent stop codon TAA in prokaryotic expression system was introduced,then the modified genes were cloned into expressing vector pBV321 and expressed in E. coli. The expressed proteins were assayed for their potencies of activity. Result:After the removal of 3′terminal non_coding region the expression level showed four times higher than that before the removal. Conclusion:3′terminal non_coding region has an inhibitory effect on its expression in E.coli,and the expression level could be increased by removing this region.
出处
《广州医学院学报》
2000年第1期15-19,共5页
Academic Journal of Guangzhou Medical College
