Thomson M, Dimmock N J
Department of Biological Sciences, University of Warwick, Coventry, United Kingdom.
Virology. 1994 Mar;199(2):366-75. doi: 10.1006/viro.1994.1133.
The polymerase chain reaction was used to amplify defective interfering (DI) Semliki Forest virus (SFV) genomes from tissue culture preparations which had in vitro interfering and in vivo mouse-protecting activity. The products were molecularly cloned and sequenced (pSFVDI-6, 2146 nt and pSFVDI-19, 1244 nt). Comparison with the sequence of the SFV genome showed that both were derived from three noncontiguous regions: the 5' terminus including the start of the nsP1 coding region, part of the nsP2 coding region, and the 3' untranslated region. Both clones possessed open reading frames, including one with the potential of encoding truncated versions of nsP1. Transcribed RNAs from pSFVDI-6 and pSFVDI-19 were translated in an in vitro system in polypeptides of M(r) 27,000 and 25,000, respectively. Although pSFVDI-6 and pSFVDI-19 did not possess the rearranged sequences or extensive repeat regions of previously cloned DI SFV genomes (Lehtovaara et al., Proc. Natl. Acad. Sci. USA 78, 5353-5357, 1981; Lehtovaara et al., J. Mol. Biol. 156, 731-748, 1982), they were derived from similar regions of the SFV genome, suggesting a common sequence requirement for DI SFV particles.
聚合酶链反应被用于从具有体外干扰和体内保护小鼠活性的组织培养制剂中扩增缺陷干扰（DI）型塞姆利基森林病毒（SFV）基因组。产物被进行分子克隆和测序（pSFVDI-6，2146个核苷酸和pSFVDI-19，1244个核苷酸）。与SFV基因组序列比较表明，两者均来源于三个不连续区域：包括nsP1编码区起始处的5'末端、nsP2编码区的一部分以及3'非翻译区。两个克隆均拥有开放阅读框，其中一个有可能编码截短形式的nsP1。来自pSFVDI-6和pSFVDI-19的转录RNA在体外系统中分别被翻译为分子量为27,000和25,000的多肽。尽管pSFVDI-6和pSFVDI-19不具备先前克隆的DI SFV基因组的重排序列或广泛重复区域（Lehtovaara等人，《美国国家科学院院刊》78，5353 - 5357，1981；Lehtovaara等人，《分子生物学杂志》156，731 - 748，1982），但它们来源于SFV基因组的相似区域，提示DI SFV颗粒存在共同的序列要求。
