Rafield L F, Knipe D M
J Virol. 1984 Mar;49(3):960-9. doi: 10.1128/JVI.49.3.960-969.1984.
The structural genes encoding the herpes simplex virus type 1 glycoprotein B and the major DNA-binding protein ICP8 have been mapped previously within the EcoRI-F restriction fragment (map coordinates 0.314 to 0.420) of the viral genome. In this study the mRNAs transcribed from these DNA sequences were identified by hybridization selection of 32P-labeled RNA and by Northern blot analysis of polyadenylated cytoplasmic RNA. A 3.4-kilobase RNA was the major mRNA homologous to the DNA sequences between coordinates 0.343 and 0.386 in which mutations in the glycoprotein B gene have been mapped. A 4.5-kilobase RNA was the major mRNA homologous to the viral DNA sequences between coordinates 0.361 and 0.417 in which mutations in the ICP8 gene have been mapped. Hybridization-selected mRNAs were translated in vitro to determine the primary translation products encoded in each region. The glycoprotein B- and ICP8-specific polypeptides were identified by immunoprecipitation with specific antisera. The translation products encoded by the glycoprotein B gene were 103,000 and 99,000 in molecular weight. The translation products encoded by the ICP8 gene were 125,000 and 122,000 in molecular weight.
编码单纯疱疹病毒1型糖蛋白B和主要DNA结合蛋白ICP8的结构基因先前已定位在病毒基因组的EcoRI - F限制片段(图谱坐标0.314至0.420)内。在本研究中,通过对32P标记RNA的杂交选择以及对多聚腺苷酸化细胞质RNA的Northern印迹分析,鉴定了从这些DNA序列转录的mRNA。一种3.4千碱基的RNA是与图谱坐标0.343至0.386之间的DNA序列同源的主要mRNA,糖蛋白B基因中的突变已定位在此区域。一种4.5千碱基的RNA是与图谱坐标0.361至0.417之间的病毒DNA序列同源的主要mRNA,ICP8基因中的突变已定位在此区域。对杂交选择的mRNA进行体外翻译,以确定每个区域编码的初级翻译产物。通过用特异性抗血清进行免疫沉淀来鉴定糖蛋白B和ICP8特异性多肽。糖蛋白B基因编码的翻译产物分子量为103,000和99,000。ICP8基因编码的翻译产物分子量为125,000和122,000。