论文部分内容阅读
目的研究人β-神经生长因子(β-NGF)cDNA在真核细胞中的表达及其表达产物的活性。方法将质粒pGEM-β-NGF进行酶切并回收β-NGF cDNA片段，构建重组真核表达载体pcDNA3-β-NGF。利用脂质体Lipofectamine方法转染COS-7细胞，对阳性克隆COS-7细胞的mRNA和培养上清分别进行Northern blot分析和观察对PC12细胞突起生长的作用。结果β-NGF基因在COS-7细胞中获得表达。阳性克隆COS-7细胞培养上清能够促进PC12细胞突起生长。结论目的基因在COS-7细胞中成功表达β-NGF蛋白，并具有良好的生物学活性。
Objective To study the expression of human β-nerve growth factor (β-NGF) cDNA in eukaryotic cells and the activity of its expressed product. Methods Plasmid pGEM-β-NGF was digested and the β-NGF cDNA fragment was recovered to construct recombinant eukaryotic expression vector pcDNA3-β-NGF. The lipofectamine method was used to transfect COS-7 cells. The mRNA and the culture supernatant of the positive clone COS-7 cells were respectively analyzed by Northern blot and observed the effect on the growth of PC12 cells. Results β-NGF gene was expressed in COS-7 cells. Positive clones COS-7 cell culture supernatant can promote PC12 cell protrusion growth. Conclusion The target gene successfully expressed β-NGF protein in COS-7 cells and has good biological activity.