Neumann G, Hobom G
Institut für Mikrobiologie und Molekularbiologie, Giessen, Germany.
J Gen Virol. 1995 Jul;76 ( Pt 7):1709-17. doi: 10.1099/0022-1317-76-7-1709.
RNA polymerase I transcription in vivo in transiently DNA-transfected cells has been used to express influenza virus vRNA molecules coding for chloramphenicol acetyltransferase (CAT) in an antisense orientation. Influenza virus superinfection provided viral RNA polymerase and other proteins required for transcriptional conversion of minus-strand vRNA into plus-strand viral mRNA molecules expressing CAT activity. This system has been used for analysis of the vRNA sequences which cooperatively constitute the vRNA promoter structure via nucleotide exchanges as well as deletions and insertions of both terminal segments. Several mutants caused greatly enhanced expression over wild-type levels, which was transmitted during serial passage of progeny virus. The data obtained for the mutations in various promoter elements support a model implicating double-stranded vRNA promoter structures in binding of viral polymerase, and in consecutive steps during initiation of RNA synthesis.
在瞬时DNA转染细胞中，利用RNA聚合酶I的体内转录来以反义方向表达编码氯霉素乙酰转移酶（CAT）的流感病毒vRNA分子。流感病毒的超感染提供了病毒RNA聚合酶以及将负链vRNA转录转化为表达CAT活性的正链病毒mRNA分子所需的其他蛋白质。该系统已用于分析通过核苷酸交换以及两个末端片段的缺失和插入共同构成vRNA启动子结构的vRNA序列。几个突变体导致表达水平比野生型大幅提高，这种提高在子代病毒的连续传代过程中得以传递。从各种启动子元件突变获得的数据支持了一个模型，该模型表明双链vRNA启动子结构参与病毒聚合酶的结合以及RNA合成起始过程中的连续步骤。
