重组ACTH(4-10)与GDNF融合蛋白及其生物活性研究

摘要

通过PCR方法构建了促肾上腺皮质激素4-10(ACTH(4-10))与胶质细胞源性神经营养因子(GDNF)的rn融合基因，并将它重组克隆到表达载体pET-28a(+)中，构建表达质粒pET-ACTH(4-10)-GDNF，转化大肠rn杆菌BL21(DE3)，经IPTG诱导可高效表达ACTH(4-10)-GDNF融合蛋白.用Ni2+-NTA树脂一步法纯化目rn的蛋白，纯度达85％以上.纯化和复性的ACTH(4-10)-GDNF融合蛋白能显著促进脊髓神经元存活，作用强rn于ACTH(4-10)及GDNF蛋白.%The chimeric gene of ACTH (4-10) with GDNF was constructed by PCR amplification. The fusedrngene was inserted into the expression vector pET-28a ( + ) and expressed in E. coli . with a level of 30% of therntotal bacterial proteins. The expressed product was purified by Ni2+ -NTA resin, up to 85% purity. The resultsrnof activity assays showed that the chimeric protein could significantly promote the survival of spinal cord neuronsrnand had a higher neurotrophic activity than ACTH (4-10) and GDNF respectively.