猪白细胞介素-6基因的非融合原核表达研究  

The study of the non-fusion prokaryotic expression of swine interleukin-6

作　　者：张桂红[1] 林海波[1] 詹国英[1] 廖明[1] 任涛[1] 罗开健[1] 曹伟胜[1] 徐成刚[1] 江经纬[1] 辛朝安[1] 

机构地区：[1]华南农业大学兽医学院禽病室,广东广州510642

出　　处：《中国预防兽医学报》2006年第4期474-477,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：广东省科技攻关项目(粤科计字【2003】292号);华南农业大学校长基金(K03005)

摘　　要：用DNA重组技术,将已克隆的猪IL-6基因阅读框片段插入非融合原核表达质粒pBV220中的适当位置,获得重组质粒pBVpIL-6,转化大肠杆菌DH5α后,42℃诱导阳性表达菌。经SDS-PAGE检测发现目的蛋白获得了高效表达,表达量约占总蛋白的20%。表达的蛋白以包涵体形式存在,包涵体经过变性溶解和复性后用酶联免疫吸附试验(ELISA)做定性定量检测。ELISA结果证实了所表达的蛋白为猪IL-6,复性后具有一定活性,并测出了复性后活性蛋白的浓度。The successful cloned open reading frame of swine IL-6 gene was inserted to the vector pBV220 of non-fusion prokaryotic expression system by DNA recombination technology. The resultant plasmid was denoted as pBVplL-6 and transformed into E. coli DH5α which was cultured at 42℃ for positive colony selection. SDS-PAGE assay indicated that the target protein was expressed in a highly efficient rate where the quantity of the expressed protein accounted for 20 % of the total expressed protein, The expressed protein existed in the inclusion body which was denaturalized, dissolved and recovered, enzyme link immunosorbent assay（ELISA） was applied for qualitative assay and quantitative assay, The result of ELISA assay indicated that after recovery the expressed swine IL-6 was bioactive, and the concentration of the recovered bioactive protein was measured,

关 键 词：猪 白细胞介素-6基因 非融合原核表达 SDS-PAGE ELISA 

分 类 号：Q78[生物学—分子生物学]