摘要
目的 :检测人 β NGF基因转染CHO细胞后培养上清中所表达的 β NGF的生物学活性及分离纯化。 方法 :采用脂质体介导的真核细胞转染 ,运用形态学观察和MTT法进行检测做定性定量分析 ,蛋白观察用SDS PAGE电泳。结果 :转染 β NGF基因的CHO细胞可分泌能促进PC12细胞生长的活性 β NGF ,且可透过血脑屏障。 结论 :转染 β NGF基因的CHO细胞所分泌的 β NGF ,接近自然合成的蛋白 ,具有较高生物学活性 ,为NGF的生物制药奠定了实验基础。
Objective:To detect the biological activity of the expression products of the human β-NGF cDNA in CHO cell line and purified the seperated protein.Methods:Human β-NGF was transfected with lipofectamine reagent into CHO cell line.The biological activity was analyzed by objection with microscope and the method of MTT.The pure protein was proved by SDS-PAGE analysis.Results:The protein in the culture supernatant of the positive CHO cells transfected with β-NGF gene could stimulate the growth of PC12 cell line and go into BBB.Conclusion:The target gene expressed successfully in the transfected CHO cell line and had good biological activity. [
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2003年第5期343-346,共4页
Chinese Journal of Immunology
基金
江苏省科委应用基金资助项目 (BJ990 70 )
