虹鳟Ⅰ型干扰素基因的表达与纯化  

Expression and Purification of Rainbow Trout(Oncorhynchus mykiss) Type Ⅰ Interferon

作　　者：张琳[1] 丁雅苓 逯茂洋[3] 夏春[3] 张秀美[1] 

机构地区：[1]山东省农业科学院畜牧兽医研究所,山东济南250100 [2]齐鲁制药有限公司,山东济南250100 [3]中国农业大学动物医学院,北京100093

出　　处：《动物医学进展》2013年第4期78-81,共4页Progress In Veterinary Medicine

摘　　要：为了获得能用于抗虹鳟(Rainbow trout,学名,Oncorhynchus mykiss)病毒性疾病生物制剂开发的高纯度Ⅰ型干扰素(rtIFN3)蛋白,克隆了虹鳟rtIFN3,构建了pET21a-rtIFN3/BL21(DE3)原核表达与发酵系,并对rtIFN3进行了表达。重组的rtIFN3分子质量约18.5ku,以包涵体形式表达,表达量占全菌蛋白量的16%。随后,提取2L发酵瓶的包涵体蛋白,经变性与复性后,再经分子筛和阴离子交换层析进行纯化。纯化后,rtIFN3SDS-PAGE电泳结果表明蛋白纯度达98%,复性率为3%。本研究获得的高纯度复性蛋白为生物制剂的研制提供了材料。In order to develop a biological agent to prevent type I interferon （rtIFN3） was obtained. Type interferon was isolated from rainbow trout in this study, then mature protein was inserted into vector pET21a which successfully expressed. The inclusion bodies extracted from 2L culture were refolded through diluted refolding method and purified with superdex 75 and Resource Q. The rtIFN3 protein was 18.5 ku and in the inclusion bodies. SDS-PAGE analysis showed that the rtlFN3 was expressed about 16 % in total proteins. 3% of the protein was correctly refolded. Successful acquisition of high-purity refolded protein was the base of biological agent development.

关 键 词：虹鳟干扰素(rtIFN) 表达 发酵 纯化 

分 类 号：S852.4[农业科学—基础兽医学]