作　　者： ; ;

机构地区： 汕头大学医学院炎症与免疫性疾病研究所

出　　处： 《免疫学杂志》 2004年第2期140-143,共4页

摘　　要： 目的　探讨一种在大肠杆菌中高效表达和简单纯化重组人白细胞介素 10 (IL 10 )的方法。方法　构建重组人IL 10表达载体pBAD Thio TOPO○R IL10 ,它编码 1个Mr3470 0融合蛋白即硫氧还蛋白 IL10。含表达质粒pBAD Thio TOPO○R IL10的工程菌经阿拉伯糖诱导、37℃培养 ,上述融合蛋白得到表达 ,表达产物经固化Ni2 + 树脂亲和层析纯化 ,用肠激酶从融合蛋白中切下靶蛋白IL 10。结果　硫氧还蛋白 IL10融合蛋白在大肠杆菌中的表达效率达 5 0 % ,并且可从细菌粗提物中一步纯化融合蛋白。 Objective To develop a method for efficient expression and rapid purification of recombinant IL-10 in E.coli. Methods Human IL-10 cDNA was PCR-amplified using gene-specific primers and cloned into a pBAD/Thio-TOPO expression vector. The pBAD/Thio-TOPO-IL10 was then used to express thioredoxin-IL10 fusion protein in E.coli at 37℃ under the induction of arabinose. The fusion protein was purified by the affinity chromatography with Ni 2+-NTA. In addition, IL-10 could be obtained after thioredoxin-IL10 was digested by enterokinase. Results The expression level of thioredoxin-IL10 fusion protein in E.coli was approximately 50%, and it can be purified by an one-step affinity chromatography. Conclusion The pBAD/Thio-TOPO expression vector provides a rapid and simple method for expression and purification of recombinant human IL-10 in E.coli.

关 键 词： 重组蛋白 表达载体 硫氧还蛋白 白细胞介素

领　　域： [生物学]