猪IL-2与IL-6的融合表达及活性研究  

Prokaryotic expression of porcine interleukin-2/interleukin-6 fusion protein

作　　者：甄洪花[1,2] 王金良[1] 单虎[2] 孙智远[2] 赵金花[2] 沈志强[1,2] 

机构地区：[1]山东省滨州畜牧兽医研究院,山东滨州256600 [2]青岛农业大学动物科技学院,山东青岛266109

出　　处：《中国预防兽医学报》2010年第10期804-807,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：山东省自主创新成果转化重大专项计划(2008ZHZX1A1103)

摘　　要：为获得具有猪白细胞介素-2(pIL-2)和猪白细胞介素-6(pIL-6)双重活性的融合蛋白,研究其作为高效免疫佐剂的可行性,本研究利用基因重组技术将克隆到的pIL-2和pIL-6的成熟肽基因利用一段柔性Linker序列串联后插入到原核表达载体pBV220中,转化大肠杆菌,42℃诱导表达得到融合蛋白pIL-6-2,对蛋白进行纯化复性后用MTT法检测其生物学活性。结果显示不同浓度pIL-6-2蛋白对小鼠脾淋巴细胞的增殖活性差异很大,0.1μg/mL浓度的pIL-6-2活性最好。本研究为利用该蛋白作为高效免疫制剂的应用奠定了良好基础。To explore the feasibility of using fusion protein of porcine interleukin-2（pIL-2） and porcine interleukin-6（pIL-6） as an immunoadjuvant,the mature peptide genes of pIL-2 and pIL-6 were linked via a hydrophilic and low charge linker sequence,and subcloned to pBV220 for prokaryotic expression.The recombinant plasmid was transformed into E.coli DH5α,BL21（DE3） and Rosetta（DE3） and then induced at 42 ℃.The expressed protein was purified and the biological activity of the protein was detected by MTT assay.The results showed that the pIL-6-2 fusion protein induced the proliferation of lymphoblast cells from the spleen of mice on a dose dependent manner,with best adjuvant effect produced at the concentration of 0.1 μg/mL.This study indicated that the pIL-6-2 protein could have the potential application as a novel efficient immunoadjuvant.

关 键 词：猪白细胞介素2 猪白细胞介素6 融合表达 生物活性 

分 类 号：S852.4[农业科学—基础兽医学]