Fas/APO-1基因真核表达质粒的构建

Construction of eukaryotic expression plasmid of Fas/APO-1 gene

目的 :构建凋亡信号受体Fas/APO - 1基因真核表达质粒 ,为进一步研究Fas/APO - 1在肿瘤细胞凋亡中的作用提供物质基础。方法 :将质粒pBluscript-FascDNA酶切后 ,回收DNA片段并提纯 ;用连接酶连接到真核表达载体pBK CMV内 ,克隆出重组基因载体pBK FascDNA。结果 :原始质粒pBluscript-FascDNA及重组质粒pBK -FascDNA酶切后 ,凝胶电泳鉴定均含有大小正确的Fas/APO - 1碱基片段。结论 :本实验成功地构建了Fas/APO - 1基因的真核表达质粒。

Objective:To construct eukaryotic express io n plasmid of Fas/APO-1 gene for further study on Fas/APO-1 function in apopt osis of cancerous cells.Methods:The plasmids pBluscr ipt-Fas cDNA were prepared by double digestion;then the DNA fragment was extract ed,purified and ligated with T4 ligase in the plasmid pBK-CMV to form pBK-Fas cD NA.Results:The original and reconstructed plasmids w ere found to contain correct-sized Fas/APO-1 cDNA fragment by agarose gel elec trophoresis after digestion.Conclusion:A eukaryotic expression plasmid of Fas/APO-1 gene was constructed successfully.