人神经营养素-4基因的原核表达与纯化  

Expression of Human Neurotrophin-4 Gene in Escherichia coliand Purification of the Product

作　　者：孙元田[1] 肖冰[1] 徐洁杰[1] 潘克俭[1] 陈平[1] 胡火珍[1] 

机构地区：[1]四川大学生命科学学院细胞生物学教研室,成都610064

出　　处：《四川大学学报（自然科学版）》2005年第3期588-591,共4页Journal of Sichuan University(Natural Science Edition)

摘　　要：以人外周血淋巴细胞基因组DNA为模板,用PCR方法扩增出神经营养素4基因(NT 4)成熟蛋白的DNA序列,并将其克隆到表达载体pET32a中,在大肠杆菌BL21(DE3)中经异丙基βD 半乳糖苷(Isopropyl beta D thiogalactopyranoside,IPTG)诱导后高效特异表达了分子量约为35kD的蛋白,诱导表达的蛋白主要存在于包涵体中,经Ni2+ NTA树脂亲和层析纯化,得到了纯度达94%的NT 4成熟蛋白的融合蛋白为进一步研究NTAmplified from peripheral blood lymphocyte by polymerase chain reaction, the neurotrophin-4 DNA sequence of mature protein was cloned on a prokaryotic pET-32a vector, and was induced to express protein in E.coli BL21(DE3). The recombination plasmid (pET-NT-4) was successfully recombined, after induced by IPTG, the recombined bacteria overexpressed a protein particularly which weighted about 35KD, and the protein presented in the formation of inclusion bodies. After purified thought Ni^(2+)-NTA affinity chromatography, the NT-4 fusion protein was obtained to a 94% purity.

关 键 词：人神经营养素-4 原核表达 融合蛋白 分离纯化 

分 类 号：Q81[生物学—生物工程]