机构地区: 中国人民解放军第一军医大学基础部全军休克微循环重点实验室
出 处: 《中国危重病急救医学》 2003年第3期163-166,共4页
摘 要: 目的 :构建人 Toll样受体 4胞浆内段 ( h TL R4C) His融合蛋白表达载体并在原核表达与纯化 ,以研究其功能及鉴定与其相互作用的蛋白。方法 :采用 PCR方法扩增 h TL R4基因编码区的胞浆内段 ,并将其重组于 p ET Dsb A2 .0载体中。重组质粒经酶切、序列鉴定分析后 ,转化大肠杆菌 BL 2 1( DE3)。结果 :用异丙基β D硫代半乳糖 ( IPTG)诱导产生 h TL R4胞浆内段的 His Dsb A融合蛋白 ,继而纯化获得了分子量约 42 kd的融合蛋白。结论 :本研究成功地构建了 h TL R4胞浆内段融合蛋白表达载体并获得高效表达 ,为进一步研究提供了重要的实验材料。 Objective:To construction of vector of histagged cytoplasmic fragment of human Toll like receptor 4 (hTLR4) and its expression in E coli.Methods:hTLR4 cytoplasmic cDNA codon domain was amplified by polymerase chain reaction(PCR) and cloned into pETDsbA2 0 plasmid expressing HisDsbA fusion protein.After being identified by the assay of restrictional enzyme and sequencing, HisDsbA fusion proteins were induced with isopropyβDthiogalactoside(IPTG) and further purified.Results:A fusion protein with molecular weight of 42 kd was obtained.Conclusion:hTLR4 which was constructed and expressed successfully under nondenaturing conditions provides a tool for further studies.
领 域: [生物学]