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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 construct a recombinant eukaryotic expression vector of human β-NGF precursor gene and lay a foundation for its expression in eukaryotic cells and obtaining the protein with nerve growth factor activity and its clinical application. Methods The recombinant plasmid pUC18-β-NGF was digested with restriction endonucleases to obtain β-NGF gene fragment and inserted into eukaryotic expression vector pcDNA3.0. The PCR products were digested with restriction endonucleases and sequenced. The inserted fragment was further identified . Results Human β-NGF gene was successfully inserted into eukaryotic expression vector pcDNA3.0. Conclusion The recombinant eukaryotic expression vector pcDNA3.0-β-NGF with human β-NGF gene is successfully constructed, which lays the foundation for the further development of NGF gene therapy for nervous system diseases.