pIRES-PML-RARα-IFN-γ重组质粒体外表达检测  

Expression of Recombination Plasmid pIRES-PML-RARα-IFN-γ in Eukaryotic Cells in Vitro

作　　者：胡刚 王旭[2,3] 李扬秋 

机构地区：[1]广东省惠东县人民医院内六科,广东惠东516300 [2]暨南大学医学院血液病研究所 [3]暨南大学再生医学教育部重点实验室

出　　处：《沈阳医学院学报》2018年第1期8-10,共3页Journal of Shenyang Medical College

基　　金：广东省医学科研基金立项资助项目(No.A2012757)

摘　　要：目的:检测pIRES-PML-RARα-IFN-γ重组质粒在真核细胞中的表达情况,为急性早幼粒细胞白血病DNA疫苗的研发提供资料。方法:将构建成功的pIRES-PML-RARα-IFN-γ重组质粒转染A549细胞株,提取总RNA,逆转录合成c DNA,采用RT-PCR法检测PML-RARα和IFN-γ基因的表达情况。提取转染后A549细胞的上清液采用Western blot和ELISA法检测PML-RARα和IFN-γ蛋白的表达情况。结果:RT-PCR结果证明转染了重组质粒的A549细胞的c DNA中含有相应的目的基因。ELISA和Western blot结果表明,重组质粒能够在A549细胞中表达出相应的目的蛋白。结论:pIRESPML-RARα-IFN-γ重组质粒能够在真核细胞中进行正常的转录及翻译。Objective: To detect the expression of recombination plasmid pIRES-PML-RARα-IFN-γ in eukaryotic cells in vitro.Methods:A549 cells were transfacted by the recombination plasmid,and then the whole RNA was extracted. The c DNA of the whole RNA was synthesized by reverse transcription,and the PML-RARα and IFN-γ gene expression were detected by RT-PCR. A549 cell supernate was extracted,and the PML-RARα and IFN-γ proteins were detected by Western blot and ELISA. Result:The results of RT-PCR proved that the c DNA of A549 cells transfected by the recombination plasmid contained the corresponding genes,and these genes could express the corresponding proteins in A549 cells. Conclusion:pIRES-PML-RARα-IFN-γ recombination plasmid with normal function has been successfully constructed.

关 键 词：PML-RARΑ IFN-Γ 早幼粒细胞白血病 DNA疫苗 

分 类 号：R733.7[医药卫生—肿瘤]