用pKKH质粒在大肠杆菌中表达HuIFN－β  

Expressing Human Fibroblast Interferon-β with Plasmid pKKH in Escherichia Colis

作　　者：许丽锋[1] 黄震[1] 白东亭[1] 蒋盘宏[1] 

机构地区：[1]卫生部北京生物制品研究所,中国药品生物制品检定所

出　　处：《微生物学免疫学进展》1996年第2期42-45,共4页Progress In Microbiology and Immunology

摘　　要：应用ＤＮＡ重组技术，将ＨｕＩＦＮ－β基因插入到质粒ｐＫＫＨ的ｔａｃ启动子下游，转化大肠杆菌ＪＭ１０１和ＪＭ１０３，经ＩＰＴＧ诱导，表达ＨｕＩＦＮ－β，收集并裂解细菌，用Ｗｉｓｈ－ＶＳＶ系统细胞病变抑制法检测生物学活性为２．１８×１０８－８．７×１０８ＩＵ／Ｌ菌液。经初步纯化ＳＤＳ－ＰＡＧＥ电泳可见分子量为２０ＫＤ较纯的表达带。Using recombinant DNA technology,HuIFM-β gene had been inserted into plasmid pKKH tac promoter downstream and transformated into Escherichia coli JM 101, JM 103 engineering bacteria and then cultured in LB culture medium, induced with IPTG and expressed rHuIFN-β.Collecting and lysising engineering bacteria,the biological activities of rHuIFN-β determinated by wish-VSV system CPE-restriction.The result was 2.18×10 8-8.7×10 8 IU/L.After primary purification, a 20 KD protein exprssing band can be seen in SDS-PAGE eletrophoretic figure.

关 键 词：重组 人成纤维细胞 干扰素 rHuIFN-β 质粒 

分 类 号：R392－33[医药卫生—免疫学]