bcl-2基因克隆、转染及其在PC12细胞中的表达

【目的】研究bcl 2基因 (bcl 2 )在PC12细胞中的表达。【方法】构建真核表达载体pcDNA3 bcl 2 ,采用脂质体介导将重组质粒导入PC12细胞 ,Westernblot和原位免疫组化检测外源基因表达。【结果】本实验成功构建了真核表达载体pcDNA3 bcl 2 ,并用脂质体介导的方法获得了稳定表达bcl 2的细胞克隆 ;Westernblot显示有 2 6ku的蛋白质表达 ,bcl 2单克隆抗体免疫组化染色呈阳性反应。【结论】重组质粒 pcDNA3 bcl 2经转染能够在PC12细胞中有效表达 ,为进一步研究 bcl 2对PC12细胞的生物学功能奠定了基础。

Study on the Expression of bcl-2 Gene in PC12 Cells by Cloning and Transfection

Objective To investigate the expression of bcl-2 gene (bcl-2) in PC12 cell line. Methods The recombinant expression plasmid pcDNA3-bcl-2 was constructed from pSFFV-bcl-2, then it was introduced into PC12 cell line by liposome method. Western blotting and immunohistochemistry in situ were applied to exam the exogenous gene expression. Results The recombinant expression plasmid pcDNA3-bcl-2 was constructed successfully. In PC12 cell line transfected by the plasmid, 26 ku protein was expressed by Western-Blot test and immunoreactivity of cells were shown by immunohistochemical staining. Conclusion bcl-2 can be expressed effectively in PC12 cell lines transfected by recombinant plasmid pcDNA3-bcl-2. The success of this research provides us with the basis for further study on the bcl-2 function in PC12 cell line.