Thompson R L, Devi-Rao G V, Stevens J G, Wagner E K
J Virol. 1985 Aug;55(2):504-8. doi: 10.1128/JVI.55.2.504-508.1985.
A herpes simplex virus type 1 (HSV-1) genetic function that is required for viral replication in the murine central nervous system was unambiguously localized. Thus, cosmid clones of either HSV-1 HindIII fragment C (0.64 to 0.87 map units) or fragment B (0.64 to 0.83 plus 0.91 to 1.0 map units) were employed to restore neurovirulence to an intertypic recombinant (RE6) that is specifically deficient in this property. The neurovirulent recombinants were generated in cell culture by cotransfecting the clone fragments and unit-length RE6 DNA and then selected in mouse brains. Either fragment efficiently conferred neurovirulence to RE6, demonstrating that no short region unique sequences are required. Analyses of the genomic structures of the neurovirulent recombinants showed that, in every case, HSV-1 information from 0.71 to 0.83 map units was incorporated into the RE6 genome. Cleavage of HindIII fragment C with EcoRI eliminated its capacity to rescue RE6. Virulence could be restored by the addition of HSV-1 BamHI fragment L (0.71 to 0.74 map units) that spans an EcoRI site at 0.72 map units. The precise location of this HSV-1 neurovirulence function is discussed.
明确确定了单纯疱疹病毒1型（HSV-1）在小鼠中枢神经系统中进行病毒复制所需的一种基因功能。因此，使用HSV-1 HindIII片段C（0.64至0.87图谱单位）或片段B（0.64至0.83加上0.91至1.0图谱单位）的黏粒克隆来恢复一种在该特性上存在特异性缺陷的型间重组体（RE6）的神经毒力。通过在细胞培养中共同转染克隆片段和单位长度的RE6 DNA，然后在小鼠脑中进行筛选，产生了具有神经毒力的重组体。任一片段都能有效地赋予RE6神经毒力，这表明不需要短的区域独特序列。对具有神经毒力的重组体的基因组结构分析表明，在每种情况下，来自0.71至0.83图谱单位的HSV-1信息都被整合到了RE6基因组中。用EcoRI切割HindIII片段C消除了其拯救RE6的能力。通过添加跨越0.72图谱单位处EcoRI位点的HSV-1 BamHI片段L（0.71至0.74图谱单位）可以恢复毒力。讨论了这种HSV-1神经毒力功能的确切位置。
