Holzer G W, Falkner F G
IMMUNO AG, Biomedical Research Center, Orth/Donau, Austria.
J Virol. 1997 Jul;71(7):4997-5002. doi: 10.1128/JVI.71.7.4997-5002.1997.
The vaccinia virus D4R open reading frame, encoding the essential DNA repair enzyme uracil DNA glycosylase, was expressed in two permanent cell lines, the rabbit kidney cell line RK13 and the human fibroblast cell line 293. The temperature-sensitive vaccinia virus mutant ts4149, which maps within D4R, was able to grow under restrictive conditions in both of these transformed cell lines. Cell clones complemented D4R function to various degrees, demonstrating complementation of an essential vaccinia virus gene by a cell line constitutively expressing the essential function. Thus, the complementing host cells allowed the rescue of a virus defective in the D4R gene, demonstrating that this system may be used for the propagation of defective cytoplasmic DNA viruses. The defective virus grew to high yields only in the engineered cell lines. The data support the hypothesis that early gene products, such as uracil DNA glycosylase, supplied in trans can fully complement essential viral functions.
编码必需的DNA修复酶尿嘧啶DNA糖基化酶的痘苗病毒D4R开放阅读框在两种永久细胞系中表达，即兔肾细胞系RK13和人成纤维细胞系293。定位在D4R内的温度敏感型痘苗病毒突变体ts4149能够在这两种转化细胞系的限制条件下生长。细胞克隆在不同程度上补充了D4R功能，证明了组成型表达必需功能的细胞系对必需痘苗病毒基因的互补作用。因此，互补宿主细胞使得D4R基因缺陷的病毒得以拯救，表明该系统可用于有缺陷的细胞质DNA病毒的增殖。有缺陷的病毒仅在工程细胞系中生长至高产量。数据支持这样的假设，即通过反式提供的早期基因产物，如尿嘧啶DNA糖基化酶，可以完全补充必需的病毒功能。
