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NT
Amplified 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.
出处
《四川大学学报（自然科学版）》
CAS
CSCD
北大核心
2005年第3期588-591,共4页
Journal of Sichuan University(Natural Science Edition)
