利用TaqMan探针技术,采用实时荧光相对定量RT-PCR的方法检测了鸡马立克氏病病毒(MDV)meq基因对鸡胚成纤维细胞(Chicken embryo fibroblasts,CEF)p53基因表达的影响,并用本室改进的TRAP法测定了端粒酶活性。结果显示,meq基因激活了CEF中原癌基因p53的表达,并且48 h的表达量是72 h的18.8倍;在对CEF和转染前后48、72 h的细胞端粒酶活性测定时也发现,端粒酶活性在转染后48 h是转染后72 h的16倍。流式细胞仪检测发现meq基因使S期细胞比例较转染前有所上升,进一步印证了meq基因是MDV中致瘤的主要因素。
Real-time relatively quantitative PCR method was used to detect p53 and chTERT gene expression in chicken embryo fibroblasts(CEFs) which was transfected by MDV meq gene,through the Taqman quantitative testing technique.At the same time,the telomerase activity was also detected by TRAP method.The result showed that meq gene activated the expression of proto-oncogene p53 in CEFs,the expression of p53 gene of 48 hrs' transfection was 10 times higher than those in 72 hrs' transfection.The telomerase activity of 48 hrs' transfection was 16 times higher than those in 72 hrs' transfection.Flow cytometry was used to distinguish cell cycle phases and it was found that meq cause the cell cycle arrested in S phase.These results further confirm that meq gene is the main oncogenic factor leading to Marek's disease lymphy cancer.