人脑源性神经生长因子(hBDNF)基因在CHO细胞的稳定表达及功能研究  

The Stable Expression of the Gene of hBDNF in CHO Cell Line and the Research of Its Function

作　　者：赵耀东[1] 杨吉成[1] 谢宇峰[1] 盛伟华[1] 张海峰[1] 苗莉[1] 缪竞诚[1] 

机构地区：[1]苏州大学细胞与分子生物学教研室,江苏苏州215123

出　　处：《苏州大学学报（医学版）》2007年第3期337-340,F0002,共5页Suzhou University Journal of Medical Science

基　　金：江苏省自然科学基金资助项目(No.BJ99070)

摘　　要：目的将人脑源性神经生长因子(hBDNF)基因转染CHO细胞后,建立稳定表达体系,并检测培养上清中所表达的hBDNF的生物学活性。方法采用脂质体介导的真核细胞转染,运用RT-PCR,Western blot及MTT法进行检测分析。结果转染hBDNF基因的CHO细胞可表达hBDNF mRNA,上清中可检出hBDNF蛋白的存在,且此上清具有促进PC12细胞生长和分化的功能。结论成功地建立了hBDNF在CHO细胞中的稳定表达体系,转染hBDNF基因的CHO细胞所分泌的hBDNF蛋白具有较高的生物学活性,为进一步研究BDNF的生物学功能和开展生物治疗奠定了实验基础。Objective To transfer the gene of human brain-derived neurotrophic factor （hBDNF） into CHO cell line and establish a stable expression system, then to detect the biologic activity of the hBDNF protein in the culture supernatant. Methods Liposome was used to mediate the transfection, RT-PCR, Westen-blot and MTT methods were used to analyze results. Results The transferred CHO cells were able to express hBDNF mRNA, and the hBDNF protein were able to be detected in the culture supernatant, and it were able to promote the growth and differentiation of PC12 cells. Conclusion We successfully establish the hBDNF gene＇s stable expression system, and the hBDNF protein secreted by the transfected CHO cells had good biologic activity, which establishes the experiment foundation for further research on BDNF＇ s biological function and launching BDNF＇ s biothera-PY

关 键 词：HBDNF 真核转染 稳定表达 功能 

分 类 号：R338[医药卫生—人体生理学]