摘要
目的:克隆大鼠胶质细胞源性神经营养因子(GDNF)全长基因,构建真核细胞表达质粒。方法:从孕17 d大鼠胚胎脑组织中提取RNA,参照《分子克隆实验指南》,合成cDNA第一、二链,加入引物PCR扩增目的基因,与pcDNA3质粒连接,构建表达质粒。结果:提取的总RNA经纯化后电泳出现18 S和28 S 2个条带,PCR扩增目的基因为630 bp的条带,测序结果为633 bp。结论:正确地克隆了大鼠GDNF基因全序列,为进一步获得重组活性的GDNF蛋白奠定基础。
Objectives: To clone rat glial cell line-derived neurotrophic factor (GDNF) total gene and to construct expression plasmid for prokaryotic cells. Methods:Total RNA was extracted from brain tissues of E17 rat embryos. According to Molecular Cloning: a Laboratory Manual, the first and second cDNA chains were synthesized and added PCR primers to amplify the target gene. The target gene was linked with pcDNA3 plasmids to construct expression vectors. Results:Purified RNA showed two bands (18S and 28S) in electrochromatography, RT-PCR product showed 630 bp band, the DNA fragment size was 633 bp. Conclusions: We have cloned the total sequence of rat GDNF correctly and laid the foundation for further obtaining GDNF protein of recombination activity.
出处
《国外医学（脑血管疾病分册）》
2004年第1期45-48,共4页
Foreign Medical Sciences Cerebrovascular Diseases
