摘要
目的 对人β-NGF前体基因的克隆及序列分析,为其在真核细胞中表达并获得具有神经生长因子活性的蛋白及临床应用打下基础。方法 从人血白细胞中提取基因组DNA,采用PCR技术,扩增出含前导肽及信号肽的β-NGF前体基因并与PUC18载体连接,经筛选得到重组质粒PUC18-β-NGF,经酶切及序列分析进行鉴定。结果 经序列测定与Genebank中已知序列(V01511)相比较,完全一致。结论 从人白细胞DNA中获得了人β-NGF前体基因,为神经生长因子的基因治疗提供了条件。
Objective To clone and analysis sequence of human gene of β- NGF gene for its expression in the mammalian cells and the clinical application.Methods By extracting the human genomic DNA from the white blood cells, the gene was cloned by PCR and PUC18 cloning method.The recombined plasmid PUC18 - β-NGF were identified by the restriction enzymes, and then the amplified cloning was sequenced and analyzed. Results The comparison of the cloned gene of β- NGF with the Genebank sequence (V01511) showed that both of the sequences were i-dentical and of 726bp length. Conclusion Cloning the β- NGF gene from the human genomic DNA had paved the way for further study on gene therapy of nervous system damage.
出处
《河南实用神经疾病杂志》
2004年第4期44-45,共2页
Henan Journal of Practical Nervous Diseases
