重组人白细胞介素10的融合表达及鉴定  

Expression and identification of recombinant human interleukin 10

作　　者：方爱平[1] 杨国成[1] 夏腊菊[1] 朱汉明[1] 

机构地区：[1]武汉大学基础医学院生物化学与分子生物学教研室,武汉430071

出　　处：《华中医学杂志》2003年第1期20-21,F004,共3页Central China Medical Journal

摘　　要：目的 研究重组人白细胞介素10(rhIL-10)载体在大肠杆菌BL21(DE3)pLyse细胞中的表达,为进一步研究IL—10在动脉粥样硬化中的作用机制奠定基础。方法 用构建成功的IL-10-PCR(?)T7/NT-TOPO(?)质粒转化大肠杆菌BL21(DE3)pLyse细胞,并通过SDS-PAGE鉴定融合表达蛋白。结果 PCR(?)T7/NT-TOPO(?)质粒载体成功载入rhIL-10基因;在异丙基硫代-β-D-半乳糖苷(IPTG)诱导下表达的蛋白质主要以包涵体形式存在。结论 在IPTG诱导下,重组的IL-10-PCR(?)T7/NT-TOPO(?)质粒载体在大肠杆菌BL21(DE3)pLyse细胞内成功表达。Objective To study the expression of recombinant human interleukin 10(IL-10) vector in E. coli BL21(DE3) pLyse cells to explore the role of IL-10 in the pathogenesis of atheroslcerosis. Methods IL-10 cDNA was subcloned into the PCRT7 /NT-TOPO bacterial expression vector. The constructed plasmids were used to transform BL21(DE3) pLyse cells. The products were conformed by 12% SDS-PAGE. Results Successfully constructed PCR T7/NT-TOPO -IL-10 was transformed into BL21(DE3) pLyse cells. Through the induction of 1 mmol/L IPTG, fusion protein was mainly expressed in the form of inclusion body. Conclusion In BL21(DE3) pLyse cells, PCR T7/NT-TOPO expression vector was successfully expressed with the induction of 1 mmol/L IPTG.

关 键 词：白细胞介素10 基因重组 大肠杆菌 基因表达 鉴定 包涵体 

分 类 号：R346[医药卫生—基础医学]