真核细胞中人神经生长因子cDNA的表达

目的 构建人神经生长因子cDNA的表达载体pcDNA3．1-NGF,并在哺乳动物细胞中进行一过性表达。为以后人神经生长因子的基因治疗打下基础。方法 在本研究室已克隆人神经生长因子cDNA的基础上。构建表达载体pcDNA3．1-NGF。用DEAE-葡聚糖转染技术将pcDNA3．1-NGF导入COS-7细胞，将人神经生长因子cDNA进行一过性表达。并采用Westernblot方法检测表达情况。以鸡胚背根神经节的生长检测表达蛋白的生物活性。结果 成功构建了表达载体pcDNA3．1-NGF，Western blot方法检测出COS-7细胞表达有目的蛋白，并具有一定的生物活性。结论 用哺乳动物细胞一过性表达了人神经生长因子，并具有一定的生物活性。

The expression of human nerve growth factor cDNA in eukaryotic cell

Objective Expression vector pcDNA3.1-NGF for human nerve growth factor(NGF) cDNA was established and was expressed tentasively in mammal cells,which is the basis of the gene therapy with human NGF. Methods Expression vector pcDNA3.1-NGF was established on the base of cloning NGF cDNA by our laboratory. pcDNA3.1-NGF was transformed into COS-7 cells using DEAE-dextraven transfection technique and expressed tentasively. The products were tested by Western blot and identified to be active by the growth of chicken embryo dorsal root nerve ganglion. Results Expression vector pcDNA3.1-NGF was established successfully,and the target protein was expressed tentasively by COS-7 cells and was of activity. Conclusion Human NGF is expressed tentasively by mammal cells and the protein is of activity.