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目的 :对引起 1980年广州登革热流行的登革 1型病毒 (GZ/ 80 )株的基因组全序列进行测定 ,为进一步了解病毒基因组结构与功能的关系 ,从分子水平探讨登革病毒的传播和流行规律及致病机制提供依据。方法 :用广州登革热流行期间从患者血清中分离的GZ/ 80株病毒感染乳鼠 ,取鼠脑制备病毒RNA。对基因组编码区分段进行RT PCR扩增 ,非编码区采用RACE法进行cDNA扩增。然后将特异产物纯化后与pGEM Teasy载体连接 ,分别进行克隆测序。结果 :GZ/ 80株基因组全长 10 735nt,5′和 3′非编码区长度分别为 94nt和 4 65nt;基因组单一的读码框架长度为 10 176nt,编码 3392个氨基酸。与登革 1型病毒 160 0 7株、WestPac74株、新加坡S2 75 / 90株和FGA/ 89株核苷酸序列的同源性分别为 94 % ,93% ,95 %和 91% ;推测的氨基酸序列的同源性分别为 98.0 % ,97.9% ,97.9%和 97.1%。结论 :GZ/ 80株基因组大小及结构与已知的登革 1型病毒其他地区分离株一致。系统发生树分析显示 ,GZ/ 80株属于第Ⅱ基因型 ,与台湾 87株、88株和泰国 80株为同一基因型。提示广州登革热流行的病原可能来自我国
OBJECTIVE: To determine the genomic sequence of dengue virus type 1 (GZ / 80) strain which caused the dengue fever in Guangzhou in 1980. To further understand the relationship between the structure and function of the virus genome, the spread and prevalence of dengue virus Laws and pathogenesis provide the basis. Methods: The neonatal rats were infected with GZ / 80 strain virus isolated from the sera of patients during the dengue epidemic in Guangzhou. Rat brain was used to prepare viral RNA. The genomic coding region was amplified by RT PCR. The non-coding region was amplified by RACE method. Then the specific product was purified and ligated into pGEM Teasy vector, respectively, for cloning and sequencing. Results: The genome of GZ / 80 strain was 10 735 nt in length and 94 nt and 4 65 nt in 5 ’and 3’ non-coding regions, respectively. The genome-wide single-length reading frame was 10 176 nt, encoding 3392 amino acids. The deduced amino acids were 94%, 93%, 95% and 91% homologous to the nucleotide sequence of dengue type 1 virus 160 7, WestPac 74, Singapore S2 75/90 and FGA / 89, respectively. The sequence homologies were 98.0%, 97.9%, 97.9% and 97.1%, respectively. Conclusion: The size and structure of GZ / 80 genome are consistent with those of other regions of dengue type 1 virus. Phylogenetic tree analysis showed that GZ / 80 strain belonged to the second genotype, which was the same genotype as 87 strains of Taiwan, 88 strains of Thailand and 80 strains of Thailand. Tip Dengue epidemic of pathogenic Guangzhou may come from China