猫干扰素基因的克隆表达及其活性测定  

Cloning and Prokaryotic Expression and Activity measurement of the IFNω Gene of feline

作　　者：许秋香[1] 刘娜[1] 刘了[1] 施振声[1] 刘文军[2] 

机构地区：[1]中国农业大学动物医学院,北京海淀100094 [2]中国科学院微生物研究所,北京海淀100080

出　　处：《中国兽医杂志》2007年第9期6-8,共3页Chinese Journal of Veterinary Medicine

摘　　要：根据已知人、马、牛、羊、兔、鹿等物种体内IFNω的基因序列分析,设计引物进行扩增。将扩增基因克隆于原核表达载体pET-His。将重组表达载体转入宿主菌进行诱导表达,表达产物以包涵体的形式存在,大小约为20kDa,表达量达到28%。将表达产物复性纯化处理后加入猫肾上皮细胞(CRFK),用水泡性口炎病毒(VSV)攻毒,测出重组IFNω的生物活性达到1.64×106U/mg,重组蛋白质的含量约为18mg/L。Based on the of the sequence of the IFNco in human,horse,cattle,sheep,rabbit,deer, and so on primers were designed. The amplified fragment was inserted into the prokaryotic to express the vector PET-His. SDS-PAGE assay showed that the target protein, with the molecular weight of 21 kDa, could be expressed in a high level and up to 28% of the total protein in E. coli cell. The target protein was added to CRFK cells, which were subsequently infected by the 100TCID50 VSV. The result showed that the activity of the recombinant IFNω was up to 1.64X 106U/mg,and the content of the recombinant IFNω was 18 mg/L.

关 键 词：猫ω干扰素基因 原核表达 重组干扰素 生物学活性 

分 类 号：Q786[生物学—分子生物学]