荣昌猪白细胞介素-2基因的原核表达  

The prokaryotic expression of Rongchang porcine IL-2

作　　者：张芳 郭万柱[2] 吴华 

机构地区：[1]中牧实业股份有限公司成都药械厂,四川成都610100 [2]四川农业大学动物生物技术中心,四川雅安625014 [3]农标普瑞纳(成都)饲料有限公司,四川成都610052

出　　处：《中国预防兽医学报》2007年第5期355-358,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家"973"项目(2004CCA01800)

摘　　要：应用DNA重组技术,将克隆的荣昌猪白细胞介素-2基因亚克隆到PET-32a(+)表达载体上,构建重组表达载体。酶切鉴定正确的重组质粒转化大肠杆菌BL21,用1 mM的HPTG诱导表达重组蛋白。RT-PCR方法检测表明白细胞介素-2基因得到了转录;对表达蛋白进行SDS-PAGE电泳,发现在相对分子质量约37 Ku处有明显的蛋白质条带,而对照组无相应条带产生。最后用MTT法检测表达蛋白的生物学活性,表明所获得的重组蛋白具有较好的生物学活性,这为利用该蛋白及其基因研制高效抗病免疫制剂和基因工程疫苗奠定了良好的基础。RIL-2 cDNA was cloned into prokaryotic expression plasmid PET-32a （＋） vector and transformed into BL21 E.coli. Protein expression was induced with 1 mM IPTG and determined by SDS-PAGE. The expressed product was about 37 Ku which existed as inclusion body. The IL-2 was purified and the biological activity was detected by MTT. The results showed that the recombinant porcine IL-2 proteins was highly effective in inducing the proliferation of porcine peripheral blood cells. This study laid foundation for research of highly efficient immune reagent and genetic vaccines.

关 键 词：荣昌猪 白细胞介素-2 表达 

分 类 号：S828.2[农业科学—畜牧学] Q786[农业科学—畜牧兽医]