Boni C, Esteban M, Pellicer A
J Gen Virol. 1984 Jul;65 ( Pt 7):1245-51. doi: 10.1099/0022-1317-65-7-1245.
Individual cloned HindIII fragments of vaccinia virus DNA were introduced into cells by DNA-mediated gene transfer. The presence of individual fragments in the different transformants was confirmed by Southern blot hybridization analysis. Several of the transformants were found to express viral sequences at various levels. The sizes of the transcripts containing vaccinia virus sequences were highly heterogeneous, with no discrete species of RNA. Positive clones contained vaccinia virus sequences in both the poly(A)+ and poly(A)- RNA fractions, although the prevalence of these sequences was variable in the two fractions. The S1 nuclease map of the 5' end of the transcripts from transformants containing the HindIII-J fragment revealed a unique 5' end, similar to RNA from virus-infected cells. In contrast, analysis of the 3' end of RNAs from these transformants showed a high degree of heterogeneity, which might explain the heterogeneity found in Northern blot patterns. In this report, it is shown for the first time that in cells transformed with vaccinia virus DNA there is proper initiation for, at least, the viral thymidine kinase gene.
通过DNA介导的基因转移，将痘苗病毒DNA的单个克隆HindIII片段导入细胞。通过Southern印迹杂交分析证实了不同转化体中单个片段的存在。发现几个转化体在不同水平上表达病毒序列。含有痘苗病毒序列的转录本大小高度不均一，没有离散的RNA种类。阳性克隆在聚腺苷酸（poly(A)+）和非聚腺苷酸（poly(A)-）RNA组分中均含有痘苗病毒序列，尽管这些序列在两个组分中的丰度不同。对含有HindIII-J片段的转化体转录本5'端的S1核酸酶图谱分析显示，其具有独特的5'端，类似于病毒感染细胞中的RNA。相反，对这些转化体RNA 3'端的分析显示高度不均一，这可能解释了Northern印迹图谱中发现的不均一性。在本报告中，首次表明在用痘苗病毒DNA转化的细胞中，至少对于病毒胸苷激酶基因存在正确的起始。
