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目的 :测定新分离登革 2型病毒福建 (FJ 10 )株 5′和 3′端非编码区序列 ,分析二级结构与功能的关系。方法 :从FJ 10株感染的乳小鼠脑中提取总RNA ,利用RACE法分别扩增 5′和 3′端cDNA片段 ,并克隆至pGEM TEasy载体中 ,然后分别测定插入片段的核苷酸序列 ,并用RNA绘图软件预测 5′和 3′端非编码区的二级结构。结果和结论 :FJ 10株 5′和 3′端非编码区长度分别为 96和 4 54个核苷酸 ,并形成复杂的茎环结构 ,可能与病毒的复制和毒力有关。
OBJECTIVE: To determine the 5 ’and 3’ non-coding regions of the newly isolated dengue 2 virus Fujian (FJ 10) strain and to analyze the relationship between secondary structure and function. METHODS: Total RNA was extracted from the brain of FJ 10 infected mice. The 5 ’and 3’ cDNA fragments were amplified by RACE and cloned into pGEM TEasy vector. The nucleotide sequence of the inserted fragment , And RNA mapping software was used to predict the secondary structure of 5 ’and 3’ non-coding regions. RESULTS AND CONCLUSION: The 5 ’and 3’ non-coding regions of FJ 10 strains were 96 and 544 nucleotides in length, respectively, forming a complex stem-loop structure, which may be related to virus replication and virulence.