摘要
目的构建人β-神经生长因子(β-NGF)前体基因重组真核表达载体,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。方法利用基因重组技术,将质粒PUC18-β-NGF进行酶切获得β-NGF基因片段,将其插入真核表达载体pcDNA3.0;用PCR技术以及酶切和测序对插入片段进行分析和进一步鉴定。结果人β-神经生长因子前体基因成功的插入真核表达载体pcDNA3.0。结论人β-神经生长因子前体基因重组真核表达载体pcDNA3.0-β-NGF构建成功,为进一步开展NGF基因治疗神经系统疾病奠定了基础。
Objective To construction the expression eukaryotic vector of human gene of β-NGF for its expression in the mammalian cells and the clinical application. Methods By gene recombination technique, human gene of β-NGF was gained from the plasmid of PUC18 -β- NGF and inserted into mammalian expression vector pcDNA3.0. The recombinant plasmid was verified with the PCR technique and restriction enzyme digcstion analysis and sequencing. Results The human gent of β- NGF was inserted into mammalian expression vector pcDN＆3.0. Conclusion The eukaryotic expression vector pcDNA3.0 -β- NGF was constructed successfully, which laid the foundation for gene therapy of nervous system diseases.
