摘要
目的探讨Neurturin(NTN)基因转染Vero细胞后的表达情况,为恒河猴帕金森病(PD)基因治疗打下基础。方法将表达载体pcDNA3-hNTN转染Vero细胞,挑选稳定表达克隆,用RT-PCR及免疫荧光分析鉴定,并对表达的人NTN蛋白进行了体外生物学活性测定。结果pcDNA3-hNTN质粒转染Vero细胞后获得了稳定表达克隆且有目的蛋白hNTN表达,表达的hNTN在体外能够促进培养的鸡胚背根神经节长出神经突起,并能促进体外培养的胚胎中脑多巴胺能神经元的存活。结论获得了能稳定表达NTN蛋白的Vero细胞株,其表达的hNTN对体外培养的中脑多巴胺能神经元具有保护作用,为移植并进行PD猴基因治疗动物实验打下了基础。
Purpose To explore the expression of Vero cells transfected by Neurturin and to lay a basis for the Parkinson＇s disease （PD） of rhesus＇s gene therapy. Methods The expression vector pcDNA3-hNTN was transfected into Vero cells. The stable expression clones were chosen and the expression of NTN gene was identified by RT-PCR and immunofluorescence. The bioactivity of NTN expressed by Vero ceils was characterized. Results The positive clones for stable expression of NTN were obtained from Veto cells which were transfected with recombinant pcDNA3-hNTN. The NTN expressed by Vero cells can stimulate the growth of nerve fibers from the dorsal root ganglia of chick embryos and promote the survival of mesencephalon dopaminergic neurons in vitro. Conclusion The Vero cells that had stable expression of hNTN were obtained and the hNTN had protective function on dopaminergic neurons, which laid a foundation for the gene therapy of Parkinson＇s disease in rhesus.
出处
《中国生化药物杂志》
CAS
CSCD
北大核心
2009年第4期221-225,共5页
Chinese Journal of Biochemical Pharmaceutics
基金
云南省科技创新强省计划基础研究重点项目(2007C0012Z)
