国家自然科学基金资助项目(30000048)及军队“十五”青年基金.
This work is supported by a grant from National Natural Sciences Foundation of China (30000048).
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,作用强于ACTH(4-10)及GDNF蛋白.
The chimeric gene of ACTH(4-10) with GDNF was constructed by PCR amplification. The fused gene was inserted into the expression vector pET-28a(+) and expressed in E.coli. with a level of 30% of the total bacterial proteins. The expressed product was purified by Ni2+-NTA resin, up to 85% purity. The results of activity assays showed that the chimeric protein could significantly promote the survival of spinal cord neurons and had a higher neurotrophic activity than ACTH(4-10) and GDNF respectively.
陈哲宇,张勇,何成,路长林,吴祥甫.重组ACTH(4-10)与GDNF融合蛋白及其生物活性研究[J].生物化学与生物物理进展,2001,28(1):67-71
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