用聚合酶链式反应(PCR)从人基因组DNA中扩增了人脑源性神经营养因子(hBDNF) cDNA和hBDNF成熟蛋白编码片段,分别克隆到pUC18中.经测序确认两个插入片段序列正确.hBDNF cDNA在CMV启动子控制下在NIH/3T3细胞中表达,用RT-PCR检测转染细胞确有BDNF mRNA存在.BDNF成熟蛋白编码序列在T7启动子控制下在E.coli中表达,SDS-PAGE表明,BDNF得到表达,以包涵体形式存在.
The cDNA of human brain-derived neurotrophic factor (hBDNF) and the fragment encoding mature hBDNF were amplified from human genomic DNA by PCR, and separately ligated into pUC18. Two inserted fragments were analysised by sequencing, which are correct.The expression of the hBDNF was controled by CMV promotor in NIH/3T3 cells transfected with recombinant plasmid, and analysed by RT-PCR, showing existance of BDNF mRNA in cells. The expression product of the mature hBDNF under the control of T7 promotor was in the inclusion body of E.coli, and visualized by SDS-PAGE analysis.
陈燕,张瑛,李金照,邓巍,夏另朝,邱蓉.人脑源性神经营养因子基因表达[J].生物化学与生物物理进展,1998,25(6):536-539
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