Meulenberg J J, de Meijer E J, Moormann R J
Central Veterinary Institute, Department of Virology, Lelystad, The Netherlands.
J Gen Virol. 1993 Aug;74 ( Pt 8):1697-701. doi: 10.1099/0022-1317-74-8-1697.
During the replication of Lelystad virus in alveolar lung macrophages, a 3'-coterminal nested set of six subgenomic RNAs (RNA2 to RNA7) is formed. These contain a common leader sequence derived from the 5' non-coding region of the genomic RNA. In this study, the sequence of the junction sites, i.e. the sites where the leader sequence joins to the body of the subgenomic RNA, was determined for all six subgenomic RNAs. For each subgenomic RNA, six to nine cDNA clones were isolated by means of reverse transcription and PCR. The nucleotide sequence at the junction site was identical for all eight cDNA clones derived from subgenomic RNA4. However, heterogeneity was observed in the nucleotide sequence surrounding the junction sites of the cDNA clones derived from subgenomic RNAs 2, 3, 5, 6 and 7. This heterogeneity suggests that the fusion of the leader to the body of the subgenomic RNA may be imprecise. The junction sites of the six subgenomic RNAs had a conserved sequence motif of six nucleotides (UCAACC or a highly similar sequence). The distance between the junction site and the translation initiation codon of the downstream open reading frame varied from nine to 83 nucleotides.
在莱利斯塔德病毒于肺泡巨噬细胞中复制期间，会形成一组3' 共末端的六个亚基因组RNA（RNA2至RNA7）嵌套序列。这些RNA包含一个源自基因组RNA 5' 非编码区的共同前导序列。在本研究中，确定了所有六个亚基因组RNA的连接位点序列，即前导序列与亚基因组RNA主体相连的位点。对于每个亚基因组RNA，通过逆转录和PCR分离出六至九个cDNA克隆。源自亚基因组RNA4的所有八个cDNA克隆在连接位点的核苷酸序列相同。然而，在源自亚基因组RNA 2、3、5、6和7的cDNA克隆连接位点周围的核苷酸序列中观察到了异质性。这种异质性表明前导序列与亚基因组RNA主体的融合可能不准确。六个亚基因组RNA的连接位点具有一个由六个核苷酸组成的保守序列基序（UCAACC或高度相似的序列）。连接位点与下游开放阅读框的翻译起始密码子之间的距离在9至83个核苷酸之间变化。
