MDV中meq基因对CEF细胞chTERT,chTR表达影响的研究

利用TaqMan探针技术,采用实时荧光相对定量RT-PCR方法检测了MDV中meq基因对鸡胚成纤维细胞(CEF)的端粒酶催化亚单位基因(chTERT)、端粒酶RNA亚基(chTR)表达水平的影响,并用TRAP法测定了端粒酶活性,用流式细胞仪检测了细胞周期的变化.实验结果表明,转染48 h后chTERT表达水平为转染后72 h的16倍,chTR表达水平在转染前后基本不变;转染48 h后CEF细胞的相对端粒酶活性是转染72 h后的12倍;在转染后72 h S期细胞的百分比较未转染细胞显著增加.

Study on the Effect of Expression Level of chTERT and chTR in Chicken Embryo Fibroblast by meq Gene

chTERT and chTR expression level in chicken embryo fibroblasts(CEF) were detected by Real-time relatively quantitative PCR method which was transfected by meq gene,through the TaqMan quantitative testing technique and the telomerase activity was detected with TRAP method and cell cycle changes also detected by flow cytometer.The results showed that the expression level of chTERT of 48 h transfection was 16 times higher than those in 72 h transfection while the expression level of chTR hadn't changes after transfection.The telomerae activity of 48 h transfection was 16 times higher than those in 72 h transfection.Flow Cytometry was used to distinguish cell cycle phases and it was found that meq cause cell cycle arrest in S phase after 72 h transfection.