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目的 :测定和分析我国登革 2型病毒福建株 (FJ 11)基因组非编码区 (NCR)序列 ,为探讨其结构特征与功能的关系提供依据。方法 :从登革 2型病毒感染的C6/36细胞中提取总RNA ,采用RACE法扩增病毒基因组 5′和 3′端片段 ,分别将其克隆至pGEM T载体 ,挑取阳性克隆进行序列测定 ;利用RNAdraw软件对非编码区二级结构进行预测。结果与结论 :我国登革 2型病毒FJ 11株基因组 5′和 3′非编码区长度分别为 96nt和 4 5 4nt,具有黄病毒共有的保守序列和二级结构。与登革 2型病毒美洲基因型比较显示 ,5′NCR第 69位核苷酸的改变对其二级结构有影响 ;3′NCR端约 70nt能形成相同的保守结构 ,而前 380nt呈现多处核苷酸的差异而导致两者预测的二级结构差别较大 ,它们可能对病毒基因组RNA的复制起重要作用
OBJECTIVE: To determine and analyze the non-coding region (NCR) sequence of China’s dengue 2 virus Fujian strain (FJ11), and to provide basis for exploring its relationship with its structure and function. METHODS: Total RNA was extracted from C6 / 36 cells infected with dengue virus type 2 (RAPD). The 5 ’and 3’ end of the viral genome was amplified by RACE and cloned into pGEM T vector. The positive clones were selected and sequenced Using RNAdraw software to predict the secondary structure of non-coding region. RESULTS AND CONCLUSION: The 5 ’and 3’ non-coding regions of dengue 2 FJ 11 genomic DNA were 96 nt and 454 nt, respectively. They shared conserved sequence and secondary structure shared by flavivirus. Compared with the dengue 2 virus American genotypes, the change of the 69th nucleotide of 5’NCR affected its secondary structure; about 70nt at the 3’NCR end formed the same conserved structure, while the first 380nt showed multiple sites Differences in nucleotides result in large differences in the predicted secondary structures, which may play an important role in the replication of viral genomic RNA