摘要
采用寡核苷酸介导的定位诱变技术,将现有的人α2a型干扰素基因中编码K23的密码子AAA定向诱变为R的密码子CGT,以构建可表达人α2b型干扰素的修饰基因。经DNA全序列分析表明,定位诱变的结果符合设计要求。在此基础上,利用原核高效表达载体pBV220在P_RP_L串联启动子的控制下在大肠杆菌中对该基因进行了表达,并对几种高效表达的人α型干扰素的抗病毒活性进行了比较。
A modified human IFN-α2b gene was generated from that of IFN-α2a by substitution of the codon AAA for lysine as amino acid residue 23 with CGT for arginine via oligonucleotide-mediated site-directed mutage nesis. The resulting gene was characterized by digestion with restriction enzymes and the mutation was confirmed by DNA sequencing of the complete coding region. The IFN-α2b gene was expressed efficiently in E. coli with a vector pBV220 under the control of bacteriophage λpRpL tandem promoters and a comparison between the antiviral activities of several subtypes and sub-subtypes of IFN-α which are all abundantly synthesized in the pBV220-E.coli system has been carried out.
出处
《病毒学报》
CAS
CSCD
北大核心
1992年第3期205-209,共5页
Chinese Journal of Virology
关键词
干扰素
修饰因基
大肠杆菌
表达
Human interferon-α2b Oligonucleotide-mediated site directed mutagensis E .coli expression system