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目的 测定登革 2型病毒 0 4株 (D2 0 4)基因组 5′和 3′末端序列。方法 从D2 0 4感染的C6 / 36细胞中提取总RNA ,以该RNA为模板 ,利用RACE法 ,分别扩增D2 0 4株的 5′和 3′末端cDNA片段。将其分别与 pGEM T载体连接得到含有 5′端 5 35bp和 3′端 5 0 3bpcDNA的重组质粒 ,并测定上述cDNA插入片段的序列。将D2 0 4的 5′和 3′端非编码区的核苷酸序列与其它登革 2型毒株进行同源性比较。结果 D2 0 4株与JAM、NGC、S1、16 6 81和PDK 5 3株的同源性分别为98 96 % ,98 96 % ,93 75 % ,98 95 % ,97 92 %和 97 72 % ,97 80 % ,90 6 5 % ,94 2 6 % ,94 2 2 %。结论 D2 0 4株除与S1株的同源性略低外 ,其余株的同源性均在 94%以上。
Objective To determine the 5 ’and 3’ end sequences of 04 (D2 0 4) genome of dengue 2 virus. METHODS: Total RNA was extracted from C6 / 36 cells infected with D2 0 4, and the 5 ’and 3’ cDNA fragments of D2 0 4 strain were amplified by RACE using this RNA as a template. This was ligated with pGEM T vector respectively to obtain a recombinant plasmid containing 5’35bp and 3’end 503bcDNA, and the sequence of the above cDNA insert was determined. The nucleotide sequences of the 5 ’and 3’ non-coding regions of D2 0 4 were compared with those of other Dengue 2 strains. Results The homologies of D2 0 4 strains to JAM, NGC, S1, 16 6 81 and PDK 5 strains were 98 96%, 98 96%, 93 75%, 98 95%, 97 92% and 97 72%, respectively. 97 80%, 90 6 5%, 94 2 6%, 94 2 2%. Conclusion D2 0 4 strains except for the homology with S1 strain is slightly lower, the rest of the strains are more than 94% homology.