Department of Biology, Georgia State University, Atlanta, GA 30302-4010, USA.
Virology. 2012 Jul 20;429(1):29-36. doi: 10.1016/j.virol.2012.04.003. Epub 2012 Apr 26.
Rubella virus (RUBV) replicons expressing a drug resistance gene and a gene of interest were used to select cell lines uniformly harboring the replicon. Replicons expressing GFP and a virus capsid protein GFP fusion (C-GFP) were compared. Vero or BHK cells transfected with either replicon survived drug selection and grew into a monolayer. However, survival was ~9-fold greater following transfection with the C-GFP-replicon than with the GFP-expressing replicon and while the C-GFP-replicon cells grew similarly to non-transfected cells, the GFP-replicon cells grew slower. Neither was due to the ability of the CP to enhance RNA synthesis but survival during drug selection was correlated with the ability of CP to inhibit apoptosis. Additionally, C-GFP-replicon cells were not cured of the replicon in the absence of drug selection. Interferon-alpha suppressed replicon RNA and protein synthesis, but did not cure the cells, explaining in part the ability of RUBV to establish persistent infections.
风疹病毒 (RUBV) 复制子表达耐药基因和感兴趣的基因，用于选择均匀携带复制子的细胞系。比较了表达 GFP 和病毒衣壳蛋白 GFP 融合蛋白 (C-GFP) 的复制子。用任一种复制子转染的 Vero 或 BHK 细胞都能在药物选择中存活并生长成单层。然而，用 C-GFP 复制子转染后的存活率比表达 GFP 的复制子高约 9 倍，虽然 C-GFP 复制子细胞的生长与未转染的细胞相似，但 GFP 复制子细胞的生长速度较慢。这都不是由于 CP 增强 RNA 合成的能力，而是与 CP 抑制细胞凋亡的能力有关。此外，在没有药物选择的情况下，C-GFP 复制子细胞不能治愈复制子。干扰素-α抑制复制子 RNA 和蛋白质的合成，但不能治愈细胞，这部分解释了 RUBV 建立持续性感染的能力。
