Isolation and characterization of a novel strain (YH01) of Micropterus salmoides rhabdovirus and expression of its glycoprotein by the baculovirus expression system.

Sun-jian Lyu; Xue-mei Yuan; Hai-qi Zhang; Wei-da Shi; Xiao-ying Hang; Li Liu; Ying-lei WU

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Isolation and characterization of a novel strain (YH01) of Micropterus salmoides rhabdovirus and expression of its glycoprotein by the baculovirus expression system.

Author: Sun-Jian LYU ¹ ; Xue-Mei YUAN ¹ ; Hai-Qi ZHANG ¹ ; Wei-da SHI ¹ ; Xiao-Ying HANG ¹ ; Li LIU ¹ ; Ying-Lei WU ¹
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1. Agriculture Ministry Key Laboratory of Healthy Freshwater Aquaculture, Key Laboratory of Fish, Health and Nutrition of Zhejiang Province, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, China.
Publication Type:Journal Article
Keywords: Micropterus salmoides; Rhabdovirus; Glycoprotein; Baculovirus system
MeSH: Animals; Baculoviridae/metabolism*; Bass/metabolism*; Carps/virology*; Cell Line; Female; Genetic Techniques; Genome, Viral; Glycoproteins/biosynthesis*; Insecta; Ovary/virology*; Phylogeny; Plasmids/metabolism*; Recombinant Proteins/biosynthesis*; Rhabdoviridae/metabolism*
From: Journal of Zhejiang University. Science. B 2019;20(9):728-739
CountryChina
Language:English
Abstract: As one of the most important aquatic fish, Micropterus salmoides suffers lethal and epidemic disease caused by rhabdovirus at the juvenile stage. In this study, a new strain of M. salmoides rhabdovirus (MSRV) was isolated from Yuhang, Zhejiang Province, China, and named MSRV-YH01. The virus infected the grass carp ovary (GCO) cell line and displayed virion particles with atypical bullet shape, 300-500 nm in length and 100-200 nm in diameter under transmission electron microscopy. The complete genome sequence of this isolate was determined to include 11 526 nucleotides and to encode five classical structural proteins. The construction of the phylogenetic tree indicated that this new isolate is clustered into the Vesiculovirus genus and most closely related to the Siniperca chuatsi rhabdovirus. To explore the potential for a vaccine against MSRV, a glycoprotein (1-458 amino acid residues) of MSRV-YH01 was successfully amplified and cloned into the plasmid pFastBac1. The high-purity recombinant bacmid-glycoprotein was obtained from DH10Bac through screening and identification. Based on polymerase chain reaction (PCR), western blot, and immunofluorescence assay, recombinant virus, including the MSRV-YH01 glycoprotein gene, was produced by transfection of SF9 cells using the pFastBac1-gE2, and then repeatedly amplified to express the glycoprotein protein. We anticipate that this recombinant bacmid system could be used to challenge the silkworm and develop a corresponding oral vaccine for fish.