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目的对云南省蚊虫病毒分离物进行分子生物学鉴定,以明确其分类地位。方法采用C6/36细胞对病毒分离物ML-13做病毒扩增,使用多种虫媒病毒基因扩增引物进行病毒基因扩增和基因序列测定。采用推导的氨基酸序列用Mega5.0软件构建种系发生树。结果 ML-13病毒可以在蚊虫细胞增殖,用OMRV和ToV-TJ病毒特异引物扩增出目的产物,序列分析显示,ML-13病毒与OMRV-AK4、ToV-TJ、OMRV-Y61、DTV、AsTV病毒衣壳蛋白和RNA依赖RNA聚合酶基因株相似性最高,分别为76%、75%、72%、67%和67%;与Toti病毒科中的DTV、ToV-TJ病毒和OMRV处于同一进化分支。结论云南省三带喙库蚊中分离的ML-13病毒为Toti病毒,这是我国首次从三带喙库蚊中分离到Toti病毒。
Objective To identify the molecular biology of mosquito virus isolates in Yunnan Province to clarify its taxonomic status. Methods C6 / 36 cells were used to amplify the virus isolate ML-13. A variety of arbovirus viral gene amplification primers were used for viral gene amplification and gene sequencing. The phylogenetic tree was constructed using the deduced amino acid sequence using Mega 5.0 software. Results The ML-13 virus could proliferate in mosquito cells. The target products were amplified by OMRV and ToV-TJ virus-specific primers. Sequence analysis showed that the ML-13 and OMRV-AK4, ToV-TJ, OMRV-Y61, DTV, AsTV The highest similarity was found between the virus capsid protein and the RNA-dependent RNA polymerase gene strains, 76%, 75%, 72%, 67% and 67%, respectively; in the same evolution as the DTV, ToV-TJ virus and OMRV in the Toti virus family Branch. Conclusion The MLT virus isolated from Culex tritaeniorhynchus in Yunnan Province is Toti virus, which is the first isolate of Toti virus from Culex tritaeniorhynchus in China.