Martinez J, St Jeor S C
J Virol. 1986 Nov;60(2):531-8. doi: 10.1128/JVI.60.2.531-538.1986.
Three virus-specific clones were isolated from a cDNA library synthesized from human cytomegalovirus (AD169)-infected cell RNA and cloned into the expression vector lambda gt11. These clones, designated C3, D10, and H10 were each found to express a human cytomegalovirus/beta-galactosidase fusion protein that was reactive with antibody prepared against purified virions. By using the cloned cDNA, we were able to identify the transcripts that code for each gene product and study the kinetics of expression during permissive infection. Our results suggest that at least two of the RNAs undergo posttranscriptional processing and appear in infected cells at immediate-early times. The authentic C3 gene product was identified by probing Western blots with antibody prepared against fusion protein fpC3.
从由人巨细胞病毒(AD169)感染的细胞RNA合成的cDNA文库中分离出三个病毒特异性克隆,并将其克隆到表达载体λgt11中。这些克隆分别命名为C3、D10和H10,每个克隆都表达一种人巨细胞病毒/β-半乳糖苷酶融合蛋白,该蛋白与针对纯化病毒粒子制备的抗体发生反应。通过使用克隆的cDNA,我们能够鉴定出编码每个基因产物的转录本,并研究允许感染期间的表达动力学。我们的结果表明,至少有两种RNA经历转录后加工,并在感染后早期出现在感染细胞中。通过用针对融合蛋白fpC3制备的抗体探测蛋白质印迹,鉴定出了真实的C3基因产物。