重组人干扰素-β在大肠杆菌中的表达与活性分析  

Expression and activity analysis of recombinant interferon-beta in E.coli

作　　者：张振龙[1] 刘建源 杨云凯 邓宛明[1] 黄颖[1] 

机构地区：[1]卫生部北京生物制品研究所 [2]北京天坛生物制品股份有限公司,北京100024

出　　处：《生物技术通讯》2001年第3期202-204,共3页Letters in Biotechnology

摘　　要：利用定点突变技术将人干扰素 β第 17位半胱氨酸编码序列突变为丝氨酸 (IFN βser17) ,DNA序列分析证明了其核苷酸序列的正确性 ,并在大肠杆菌TAP10 6中获得高效表达 ,IFN βser17表达量达 2 0 % ,用Westernblot得到单一条带 ,细胞病变抑制法测定其比活性达 (2～ 3)× 10 7U mg。INF βser17的比活性和稳定性均超过天然人IFN β。In the natural interferon-beta, the residue Cys at the position 17 was replaced with Ser by sits-specific mutagenesis. Analysis of DNA sequencing demonstrated that IFN-βser17 has the high expression in E.coli TAP106, the expression level was more than 20%. The result of Western blot showed that the expression protein was correct. By the way of WISH-VSV cell inhibition, relative activity was about (2～3)×10 7 U/mg. The activity and stability of INF-βser17 were better than the natural interferon-beta.

关 键 词：重组人干扰素-β 点突变 基因表达 大肠杆菌 活性分析 

分 类 号：R394.8[医药卫生—医学遗传学] TQ460.38[医药卫生—基础医学]