Endoh D, Ikegawa S, Kon Y, Hayashi M, Sato F
Laboratory of Radiation Biology, Hokkaido University, Sapporo.
Jpn J Vet Res. 1995 Dec;43(3-4):109-24.
The ICP4 homolog of Marek's disease virus (MDV ICP4) is a possible candidate for the transactivator of the early genes. We transfected MDCC-MSB-1 (MSB-1) tumor cells with plasmid including a coding region of MDV ICP4 using cationic liposome. As carriers for intranuclear transport, high mobility group -1 and -2 proteins were bound to the plasmid DNA before forming liposomes. We detected transcripts from the plasmid 2 hr after transfection by quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) analysis. We also detected abundant transcripts of endogenous ICP4 2-96 hr after transfection. These data suggested that expression of introduced MDV ICP4 gene enhanced the expression of endogenous MDV ICP4. On the other hand, quantitative PCR analysis for virus genome DNA indicated no significant alteration of copy number of virus genome in transfected MSB-1 cells, suggesting that reactivation of virus requires more than turning on MDV ICP4 gene.
马立克氏病病毒的ICP4同源物(MDV ICP4)可能是早期基因反式激活因子的一个候选物。我们使用阳离子脂质体将包含MDV ICP4编码区的质粒转染到MDCC-MSB-1(MSB-1)肿瘤细胞中。作为核内运输的载体,高迁移率族-1和-2蛋白在形成脂质体之前与质粒DNA结合。通过定量逆转录聚合酶链反应(RT-PCR)分析,我们在转染后2小时检测到了来自质粒的转录本。我们还在转染后2-96小时检测到了内源性ICP4的大量转录本。这些数据表明,导入的MDV ICP4基因的表达增强了内源性MDV ICP4的表达。另一方面,对病毒基因组DNA的定量PCR分析表明,转染的MSB-1细胞中病毒基因组拷贝数没有显著变化,这表明病毒的重新激活需要的不仅仅是开启MDV ICP4基因。