Preston C M, McGeoch D J
J Virol. 1981 May;38(2):593-605. doi: 10.1128/JVI.38.2.593-605.1981.
mRNA's homologous to the herpes simplex virus type 1 DNA restriction endonuclease fragment BamHI p, which contains the thymidine kinase gene, have been identified and mapped by hybrid-arrested translation and mRNA selection. Such mRNA's, when translated in vitro, directed the synthesis of polypeptides of apparent molecular weights 43,000 (VI43) and 39,000 (VI39). mRNA for enzymatically active thymidine kinase was enriched by more than 20-fold after selection. Mapping was carried out with restriction endonuclease fragments of BamHI p, and locations of the 5' and 3' termini of VI43 mRNA were deduced. Analysis of nucleotide sequences around the 5' terminus revealed several consensus sequences commonly found at the start of eucaryotic mRNA's and which are presumably involved in initiation of transcription by RNA polymerase II. Translation of mRNA's for VI43, VI39, and the thymidine kinase enzyme was arrested only by a 1,170-base-pair region of BamHI p. Since this region is insufficient for adjacent genes, coding sequences for VI43 and VI39 must overlap; the possible relationship of these two polypeptides is discussed. A virus-induced product equivalent to VI39 was detected in infected cells.
已通过杂交捕获翻译和mRNA选择鉴定并定位了与单纯疱疹病毒1型DNA限制性内切酶片段BamHI p同源的mRNA，该片段包含胸苷激酶基因。此类mRNA在体外翻译时，指导合成表观分子量为43,000（VI43）和39,000（VI39）的多肽。选择后，具有酶活性的胸苷激酶的mRNA富集了20多倍。用BamHI p的限制性内切酶片段进行定位，并推导了VI43 mRNA的5'和3'末端位置。对5'末端周围核苷酸序列的分析揭示了在真核mRNA起始处常见的几个共有序列，推测这些序列参与RNA聚合酶II的转录起始。VI43、VI39和胸苷激酶酶的mRNA翻译仅被BamHI p的1170个碱基对区域阻断。由于该区域对相邻基因而言不足，VI43和VI39的编码序列必定重叠；讨论了这两种多肽的可能关系。在感染细胞中检测到一种相当于VI39的病毒诱导产物。
