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从人胎脑组织中提取总ＲＮＡ，以ＲＴＰＣＲ方法获取编码胶质细胞源神经营养因子（ＧＤＮＦ）成熟蛋白的ｃＤＮＡ。将人ＧＤＮＦｃＤＮＡ插入含Ｔ７启动子的质粒ｐＥＴ２８ａ（＋），构建表达质粒ｐＥＴＧＤＮＦ，转化大肠杆菌获得表达菌株ＢＬＧＤＮＦ，经诱导表达的ＧＤＮＦ形成包含体。凝胶自动扫描分析表明，表达量约占菌体总蛋白的３０％以上。用纯化的ＧＤＮＦ蛋白免疫新西兰兔制备了ＧＤＮＦ抗血清。纯化和复性的ＧＤＮＦ蛋白能显著促进多巴胺能神经元的存活。
Total RNA was extracted from human fetal brain tissue and cDNA encoding the mature protein of glial cell line-derived neurotrophic factor (GDNF) was obtained by RT-PCR. The human GDNF cDNA was inserted into the plasmid pET28a (+) containing the T7 promoter to construct the expression plasmid pETGDNF, and transformed into E. coli to obtain the expression strain BLGDNF. The induced expression of GDNF formed inclusion bodies. Gel automated scanning analysis showed that the expression amount of about 30% of total bacterial protein. GDNF antiserum was prepared by immunizing New Zealand rabbits with purified GDNF protein. Purified and renatured GDNF protein can significantly promote the survival of dopaminergic neurons.