人α2b型干扰素基因的修饰合成及其在大肠杆菌中的表达  

GENERATION OF A MODIFIED INTERFERON-α2b GENE AND ITS EXPRESSION IN ESCHERICHIA COLII

作　　者：曾庆[1] 金冬雁[1] 金奇[1] 胡钢[1] 周圆[1] 李玉英[1] 王珣章[2] 侯云德[1] 

机构地区：[1]中国预防医学科学院病毒学研究所病毒基因工程国家重点实验室 [2]中山大学昆虫学研究所

出　　处：《病毒学报》1992年第3期205-209,共5页Chinese Journal of Virology

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.

关 键 词：干扰素 修饰因基 大肠杆菌 表达 

分 类 号：R394.8[医药卫生—医学遗传学]