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目的 对新近分离的导致 1999年福建省登革热流行的登革 2型病毒FJ 10株进行基因组全序列测定及系统发生树分析。方法 利用RT PCR和 5′、3′RACE法扩增FJ 10株cDNA ,并进行克隆测序。利用DNASTAR软件的Clustal方法绘制系统发生树。结果 FJ 10株基因组全长 10 72 3个核苷酸 ,有 1个单一开放读码框架 (ORF ,第 97～ 10 2 6 9nt) ,编码 3391个氨基酸 ,5′和 3′非编码区长度分别为 96和 45 4个核苷酸。通过与标准株NGC株和我国其他地区分离株DEN2 0 4、43、44株比较 ,核苷酸同源性分别为 94.0 %、92 .8%、93.9%和 93.9% ,氨基酸同源性分别为 97.9%、97.2 %、97.7%和97.9%。以 47株登革 2型病毒E/NS1连接区 2 40个核苷酸序列进行系统发生树分析 ,福建株与印度尼西亚和斯里兰卡分离株亲缘关系较近 ,同属于第Ⅳ基因型。结论 FJ 10株基因组全序列一级结构与其他登革 2型病毒类似 ,其基因型不同于我国其他地区分离株DEN2 0 4、43和 44株。
Objective To analyze the genome-wide sequence and phylogenetic tree of the recently isolated FJ 10 strain of dengue 2 virus that caused the dengue epidemic in Fujian Province in 1999. Methods FJ 10 cDNA was amplified by RT PCR and 5 ’, 3’RACE and cloned and sequenced. Phylogenetic trees were drawn using the Clustal method of DNASTAR software. Results The genome of FJ was 10 723 nucleotides in length and contained a single open reading frame (ORF, 97-102 6 9nt) encoding 3391 amino acids with 5 ’and 3’ non-coding regions 96 and 45 4 nucleotides. Nucleotide homology was 94.0%, 92.8%, 93.9% and 93.9%, respectively, compared with NGC strain of standard strain and DEN2 0, 443, 44 strains of other regions in China. The amino acid homologies were 97.9%, 97.2%, 97.7% and 97.9%. Phylogenetic analysis of phylogenetic tree was performed using 40 nucleotide sequences of 47 Dengue 2 virus E / NS1 junctional regions. Fujian strain was closely related to Indonesia and Sri Lanka isolates, belonging to the fourth genotype. CONCLUSION: The primary structure of FJ 10 isolates is similar to other Dengue 2 viruses. The genotypes of FJ 10 isolates are different from those of other isolates from China.