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用逆转录-聚合酶链反应(RT-PCR)以大鼠纹状体总RNA为模板,扩增了大鼠胶质细胞源性神经营养因子(rGDNF)成熟序列的cDNA片段,将此cDNA克隆到载体pGEM-T中,对重组质粒进行限制酶切分析和序列测定,确定为含rGDNFcDNA的重组质粒,将该rGDNFcDNA重组到融合表达载体pGEX—1λT内,在大肠杆菌中获得了较高的表达。
The cDNA fragment of rat glial cell line-derived neurotrophic factor (rGDNF) mature sequence was amplified by reverse transcription-polymerase chain reaction (RT-PCR) using total RNA of rat striatum as a template. The cDNA clone To pGEM-T vector, restriction analysis and sequence analysis of the recombinant plasmid were carried out to confirm that it was a recombinant plasmid containing rGDNF cDNA. The rGDNF cDNA was recombined into the fusion expression vector pGEX-1λT to obtain higher expression in E. coli .