Collins P L, Dickens L E, Buckler-White A, Olmsted R A, Spriggs M K, Camargo E, Coelingh K V
Proc Natl Acad Sci U S A. 1986 Jul;83(13):4594-8. doi: 10.1073/pnas.83.13.4594.
Complete sequences for the intergenic regions of the genome of human respiratory syncytial virus were obtained by dideoxynucleotide sequencing using synthetic oligonucleotides. These experiments established that the 10 respiratory syncytial viral genes are arranged, without additional intervening genes, in the order 3' 1C-1B-N-P-M-1A-G-F-22K-L 5'. For the first nine genes, the exact gene boundaries were identified by comparison of the genomic sequences with previously determined mRNA sequences. The intergenic regions varied in length from 1 to 52 nucleotides and lacked any obvious conserved features of primary or secondary structure except that each sequence ended (3' to 5') with an adenosine residue. The exact start site of the 10th gene, the L gene, was not determined. However, RNA blot hybridization using a synthetic oligonucleotide designed from the genomic sequence mapped the L gene to within 54 nucleotides of the end of the penultimate 22K gene. The lack of conservation of chain length and nucleotide sequence for the respiratory syncytial viral intergenic regions, together with the complexity of the genetic map, contrasts with previous observations for other nonsegmented negative-strand viruses.
通过使用合成寡核苷酸进行双脱氧核苷酸测序，获得了人类呼吸道合胞病毒基因组基因间区域的完整序列。这些实验证实，10个呼吸道合胞病毒基因按3' 1C-1B-N-P-M-1A-G-F-22K-L 5'的顺序排列，其间没有其他插入基因。对于前九个基因，通过将基因组序列与先前确定的mRNA序列进行比较，确定了精确的基因边界。基因间区域的长度从1到52个核苷酸不等，除了每个序列（3'至5'）末端都有一个腺苷残基外，缺乏任何明显的一级或二级结构保守特征。第10个基因即L基因的确切起始位点尚未确定。然而，使用根据基因组序列设计的合成寡核苷酸进行的RNA印迹杂交将L基因定位在倒数第二个22K基因末端的54个核苷酸范围内。呼吸道合胞病毒基因间区域链长度和核苷酸序列缺乏保守性，以及遗传图谱的复杂性，与之前对其他非节段负链病毒的观察结果形成对比。
