摘要
用已构建好的重组表达载体pGEX-4T-IL-4转染E.coli,在最适条件下诱导表达,表达产物经SDS-PAGE分析,表达出38 ku的融合蛋白,表达量占菌体总蛋白的25%,表达产物主要以包涵体的形式存在;对表达产物经变性、复性及初步纯化后再结合饱和硫酸铵沉淀和Sephadex-G100凝胶层析柱进一步纯化,所得蛋白的纯度约在90%以上。在体外利用MTT法检测其生物学特性,结果表明其具有较好的生物学活性,本试验为获得猪IL-4重组蛋白奠定了基础。
The recombinant expression vector pGEX-IL-4 was induced by IPTG in optimal conditions in E. coll. SDS-PAGE analysis showed that the expressed fusion protein with a molecular weight of 38 ku existed in the inclusion body. The fusion protein, which was of 25% in total bacterial proteins, was denatured, renatured, and purified with saturated ammonium sulfate precipitation and Sephadex-G100 with the purity of above 90%. MTT analysis indicated that purified protein has good bioactivity, which has laid a foundation for the production and development of recombinant protein of porcine IL-4.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2007年第10期1072-1076,共5页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家高技术研究发展计划(863)项目(2006AA10A203)
兰州市科技攻关项目(05-1-47)
关键词
猪白细胞介素4
重组表达
纯化
活性分析
porcine interleukin 4
recombinant expression
purification
bioactivity analysis
