Departamento de Medicina Preventiva, Salud Pública y Microbiología, Facultad de Medicina, Universidad Autónoma de Madrid, Arzobispo Morcillo 4, 28029 Madrid, Spain.
Arch Virol. 2010 Apr;155(4):515-23. doi: 10.1007/s00705-010-0613-4. Epub 2010 Feb 27.
The pseudorabies virus (PRV) glycoprotein known as gG is generally regarded as an early protein, and the immediate early IE180 protein regulates its expression during infection. This study, however, provides evidence that although induction by IE180 is observed, the expression of a marker protein (EGFP), or gG itself, under the control of the gG promoter, can also occur independently of the expression of IE180. This result was demonstrated both with transient transfection assays using plasmids and with viral infections. In transient transfections, the expression under control of the gG promoter depends on the cell type and surprisingly, can be 1.3-fold higher than the expression under the control of the IE180 promoter in Hela Tet-Off cells. Recombinant PRV S3 was constructed by replacing gE in the PRV genome with a chimeric transgene, expressing EGFP under the control of the gG promoter. In PK15 cells infected with NIA-3 wild-type virus or with S3 recombinant virus, expression of gG PRV mRNA (or EGFP mRNA) under the control of the gG promoter in the presence of cycloheximide was detected by RT-PCR. This again indicates that some basal expression was produced in infected cells independently of IE180. This expression was augmented by IE180 protein in both plasmid transfections and viral infections.
伪狂犬病毒(PRV)糖蛋白 gG 通常被认为是一种早期蛋白,而即刻早期 IE180 蛋白在感染过程中调节其表达。然而,本研究提供的证据表明,尽管观察到 IE180 的诱导,但在 gG 启动子控制下表达标记蛋白(EGFP)或 gG 本身也可以独立于 IE180 的表达发生。这一结果通过使用质粒的瞬时转染测定和病毒感染来证明。在瞬时转染中,gG 启动子控制下的表达依赖于细胞类型,令人惊讶的是,在 Hela Tet-Off 细胞中,其表达可以比 IE180 启动子控制下的表达高 1.3 倍。通过用在 gG 启动子控制下表达 EGFP 的嵌合转基因替换 PRV 基因组中的 gE,构建了重组 PRV S3。在感染 NIA-3 野生型病毒或 S3 重组病毒的 PK15 细胞中,通过 RT-PCR 检测到在存在环己酰亚胺的情况下,gG PRV mRNA(或 EGFP mRNA)在 gG 启动子控制下的表达。这再次表明感染细胞中独立于 IE180 产生了一些基础表达。这种表达在质粒转染和病毒感染中均被 IE180 蛋白增强。