Wittek R, Hänggi M, Hiller G
J Virol. 1984 Feb;49(2):371-8. doi: 10.1128/JVI.49.2.371-378.1984.
Cell-free translation of total RNA isolated from vaccinia virus-infected cells late in infection results in a complex mixture of polypeptides. A monospecific antibody directed against one of the major structural proteins of the virus particle immunoprecipitated a single polypeptide with a molecular weight of 11,000 (11K) from this mixture. Immunoprecipitation was therefore used to identify the structural polypeptide among the in vitro translation products of RNA purified by hybridization selection to restriction fragments of the vaccinia virus genome. This allowed us to map the mRNA coding for the 11K polypeptide to the extreme left-hand end of the HindIII E fragment. Detailed transcriptional mapping of this region of the genome by nuclease S1 analysis revealed the presence of a late RNA transcribed from the rightward-reading strand. Its 5' end mapped at ca. 130 base pairs to the left of the HindIII site at the junction between the HindIII F and E fragments. The map position of this RNA coincided precisely with the map position of the late message coding for the 11K polypeptide.
从感染后期的痘苗病毒感染细胞中分离出的总RNA进行无细胞翻译，会产生复杂的多肽混合物。一种针对病毒颗粒主要结构蛋白之一的单特异性抗体，从该混合物中免疫沉淀出一种分子量为11,000（11K）的单一多肽。因此，免疫沉淀被用于在通过与痘苗病毒基因组的限制性片段进行杂交选择纯化的RNA的体外翻译产物中鉴定结构多肽。这使我们能够将编码11K多肽的mRNA定位到HindIII E片段的最左端。通过核酸酶S1分析对基因组的该区域进行详细的转录图谱分析，揭示了从右向阅读链转录的晚期RNA的存在。其5'端位于HindIII F和E片段交界处的HindIII位点左侧约130个碱基对处。这种RNA的图谱位置与编码11K多肽的晚期信使RNA的图谱位置精确重合。
