新型融合基因hIFNα1-CB在大肠杆菌中表达  

Expression of a Novel Fusion Gene hIFN α1-CB in E. coli

作　　者：邱定红[1] 毛晓华[1] 高向东[2] 李新荣[1] 林碧蓉[2] 徐蘅 

机构地区：[1]东南大学基础医学院,南京210009 [2]中国药科大学生物制药学院,南京210009

出　　处：《中国药科大学学报》2002年第6期521-525,共5页Journal of China Pharmaceutical University

基　　金：铁道部科技基金;东南大学教育基金资助项目 ( JP72 0 13;92 2 30 0 1165 )

摘　　要：目的　构建人干扰素 α1和抗菌肽 B的融合蛋白表达质粒 p HC,并初步研究融合蛋白的抗病毒活性。方法　重组质粒 p HC转化 E.coli JM83 ,温控诱导表达 ,SDS- PAGE制备蛋白质 ,病变抑制法测定其抗病毒比活性。结果　得到分子量约为 2 1ku的目的蛋白 ,抗病毒比活性为 2 .62× 10 6 IU/ m g。结论　成功构建了融合蛋白表达质粒 p HC,重组蛋白的构象及 C-端的AIM To construct a plasmid expressing fusion protein,and to study the antiviral activity of the proteins.METHODS Artifically modified hIFNα1 cDNA was fused precisely to the reading frame of CB cDNA and further cloned into expression vector pBV220. The plasmid was transformed into E. coli and expressed under thermal induction. The fusion protein was purified by preparetive SDS-PAGE.RESULTS Expression analysis revealed accounts for 2.8% of total cellular proteins. Antiviral activity of this fusion protein was 2.62×10 6 IU/mg by CPE. CONCLUSION We succeeded in constructing the recombinant plasmid expressing hIFNα1-CB fusion proteins. Factors such as non-native conformation and β-amidation were responsible for the low activity of hIFNα1-CB fusion molecule.

关 键 词：大肠杆菌 表达 基因重组 人干扰素 抗菌肽B 抗病毒活性 

分 类 号：Q786[生物学—分子生物学]