Carmichael E P, Kosovsky M J, Weller S K
Department of Microbiology, University of Connecticut Health Center, Farmington 06032-9984.
J Virol. 1988 Jan;62(1):91-9. doi: 10.1128/JVI.62.1.91-99.1988.
Cell lines were generated by cotransfection of Vero cells with pSV2neo and a plasmid containing the herpes simplex virus type 1 (HSV-1) EcoRI D fragment (coordinates 0.086 to 0.194). One such cell line (S22) contained the genes for alkaline exonuclease and several uncharacterized functions. Three mutant isolates of HSV-1 strain KOS which grew on S22 cells but not on normal Vero cells were isolated and characterized. All three mutants (hr27, hr48, and hr156) were defective in the synthesis of viral DNA and late proteins when grown in nonpermissive Vero cells. Early gene expression in cells infected with these host range mutants appeared to be normal at the nonpermissive condition. The mutations were mapped by marker rescue to a 1.5-kilobase fragment (coordinates 0.145 to 0.155). The mutation of one of these mutants, hr27, was more finely mapped to an 800-base-pair region (coordinates 0.145 to 0.151). This position of these mutations is consistent with the map location of a putative 94-kilodalton polypeptide as determined by sequence analysis (D. McGeoch, personal communication). Complementation studies demonstrated that these mutants formed a new complementation group, designated 1-36. The results presented in this report indicate that the 94-kilodalton gene product affected by these mutations may have a direct role in viral DNA synthesis.
通过用pSV2neo和含有单纯疱疹病毒1型（HSV-1）EcoRI D片段（坐标0.086至0.194）的质粒共转染Vero细胞来生成细胞系。一个这样的细胞系（S22）含有碱性核酸外切酶基因和几种未鉴定的功能。分离并鉴定了HSV-1 KOS株的三个突变分离株，它们能在S22细胞上生长但不能在正常Vero细胞上生长。当在非允许性Vero细胞中生长时，所有三个突变体（hr27、hr48和hr156）在病毒DNA和晚期蛋白的合成方面存在缺陷。在非允许条件下，感染这些宿主范围突变体的细胞中的早期基因表达似乎正常。通过标记拯救将突变定位到一个1.5千碱基的片段（坐标0.145至0.155）。其中一个突变体hr27的突变被更精细地定位到一个800碱基对的区域（坐标0.145至0.151）。这些突变的位置与通过序列分析确定的假定94千道尔顿多肽的图谱位置一致（D. McGeoch，个人交流）。互补研究表明，这些突变体形成了一个新的互补组，命名为1-36。本报告中的结果表明，受这些突变影响的94千道尔顿基因产物可能在病毒DNA合成中具有直接作用。
