重组人IL-11的原核表达、纯化和鉴定  

Prokaryotic Expression,Purification,and Identification of Recombinant Human IL-11 Protein

作　　者：唐佳[1] 许晓玲[1] 聂小霞[1] 茅奇峰[1] 高基民[1] 

机构地区：[1]温州医学院浙江省模式生物技术与应用重点实验室,温州325035

出　　处：《生物医学工程学杂志》2012年第3期530-533,共4页Journal of Biomedical Engineering

基　　金：浙江省自然科学基金(杰出青年团队)资助项目(R2080407)

摘　　要：以本实验室构建的pET24a-hrIL-11表达载体为模板,PCR扩增重组人白细胞介素11(hrIL-11)的基因,克隆入pET21a表达载体,转化BL-21(DE3),经异丙基-β-D-硫代半乳糖苷(IPTG)诱导产生hrIL-11/His融合蛋白,并经Western blot实验确认。原核表达的hrIL-11/His融合蛋白经变性后,利用亲和层析纯化,目的蛋白纯度达95%以上。纯化后蛋白经尿素梯度复性后,比活达到1×106 IU/mg。研究结果为进一步研究该蛋白在促进血小板增殖等方面的作用奠定了基础。A DNA fragment encoding recombinant human interleukin 11 （hrlL-11） was obtained by PCR from previ- ously constructed pET24a-hrlL-11 plasmid. Then pET21a-hrlL-11 expression vector was constructed routinely and transformed into BL-2I （DE3）. By the induction of Isopropyl-l-thio-β-Dgalactoside （IPTG）, hrlL-11 protein was highly expressed at about 200/00 of the total bacterial proteins and was identified by Western blot. After purification with Ni-NTA affinity chromatography and refolding with renaturation buffer, the purity of the target hrlL-11 protein reached 95% and its biology activity was 1×10^6IU/mg, determined by stimulating the proliferation of Tl165, which facilitates further researches into effects of IL-11 on platelet proliferation and other function

关 键 词：重组人白细胞介素11 原核表达 纯化 生物学活性 

分 类 号：R392[医药卫生—免疫学]