摘要
本文合理设计新型重组人干扰素α2b(rh-INFα2b)基因序列,采用组合突变策略,对第52-53-55、103-107和121-125位氨基酸残基定向突变。将突变基因连接于原核表达载体pET28a上进行诱导表达,所获得的新型rh-IFNα2b的抗病毒活性效价达9.3×107IU/mg,比对照样品(1×106IU/mg)高93倍。结果表明:所采用邻近多点突变方法,有效组合了位点效应,大幅度提高了干扰素的生物活性,获得高抗病毒活性的rh-IFNα2b。
In order to create a novel recombinant human interferon α2b （rh-IFN α2b） with higher biological activity,we subjected the rational designed sequence of rh-IFN α2b to direct evolution by strategy of the combinatorial mutagenesis.The amino acid residues at multiple sites of 52-53-55,103-107,and 121-125 were simultaneously mutated.The resulted gene of the mutated rh-IFN α2b was cloned into the pET28a and expressed in E.coli BL21 Condon plus（RIL）.The anti-virus activity of the novel interferon α2b was 9.3×107 IU/mg,93 times higher than the wild type（1×106 IU/mg）.The results showed that the multiple point mutation used in this study could effectively combine the site effects of rh-IFN α2b and increase its biological activity.
出处
《生物医学工程学杂志》
EI
CAS
CSCD
北大核心
2011年第2期347-351,共5页
Journal of Biomedical Engineering
基金
天津市自然科学基金资助项目(06YFJMC07300)
高等学校博士学科点专项科研基金资助项目(20090032110015)
关键词
重组人干扰素
组合突变
大引物扩增
抗病毒活性
Recombinant human interferon（rh-IFN）
Combinatorial mutagenesis
Megaprimer amplification
Anti-virus activity
