1株库蠓源版纳病毒的分离与全基因组序列分析

doi:10.16431/j.cnki.1671-7236.2021.03.001

Abstract

Abstract: To investigate the variety and genetic characteristics of arboviruses epidemic in Yunnan province,Culicoides were collected in Shizong county and inoculated cells for virus isolation and identification.Whole genome sequence of the isolated virus was obtained by full-length cDNA amplification (FLAC) and next generation sequencing (NGS),then analyzed through sequence alignment and phylogenetic tree.The results showed that a virus strain (YNSZ043) causing cytopathic effect on C6/36 cells was isolated from collected Culicoides specimens.The genome of the isolated virus was composed with double-stranded segmented RNA,showing migration pattern of "2-4-3" by agarose gel electrophoresis.The viral particle of the isolated virus was about 70 to 80 nm in diameter exhibiting morphological feature of "finger ring",with fibrous protrusions on the surface.The genome size of the YNSZ043 was 20 683 bp in length with 12 gene segments ranging from 3 762 bp (Seg-1) to 861 bp (Seg-12),with nucleic acid sequence identities ranging from 64.8% to 99.6% and amino acid sequence identities ranging from 58.8% to 100%,compared with other Chinese BAV strains.On the phylogenetic tree,YNSZ043 clustered together with other Chinese BAV strains forming an independent Chinese evolutionary branch.YNSZ043 belonged to A2 genotype by phylogenetic analysis of Seg-12 which decided the genotype of BAV.Maximum nucleic and amino acid sequence identity of Seg-5/VP5 were observed between YNSZ043 and BAV strains isolated from Vietnam,which were 97.1% and 97.6%,respectively,indicated there might be Seg-5 reassortment between Vietnamese strains and YNSZ043 strain.The present study enriched the genome sequence of BAV,provided a foundation for further epidemiological studies of BAV in Yunnan.

Key words: Culicoides; Banna virus(BAV); isolation and identity; whole-genome sequence; genome reassortment

CLC Number:

S852.65

LI Zhanhong, ZHU Jianbo, KOU Meiling, YANG Zhenxing, LI Zhuoran, NIU Baosheng, YAO Pingfen, LI Huachun, YANG Heng. Isolation and Whole Genome Sequence Analysis of a Banna Virus Strain from Culicoides[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(3): 781-791.

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Isolation and Whole Genome Sequence Analysis of a Banna Virus Strain from Culicoides

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