title: Construction of green fluorescent protein-tagged recombinant iridovirus to assess viral replication.
abstract: Green fluorescent protein-tagged recombinant virus has been successfully applied to observing the infective dynamics and evaluating viral replication. Here, we identified soft-shelled turtle iridovirus (STIV) ORF55 as an envelope protein (VP55), and developed a recombinant STIV expressing an enhanced green fluorescent protein (EGFP) fused to VP55 (EGFP-STIV). Recombinant EGFP-STIV shared similar single-step growth curves and ultrastructural morphology with wild type STIV (wt-STIV). The green fluorescence distribution during EGFP-STIV infection was consistent with the intracellular distribution of VP55 which was mostly co-localized with virus assembly sites. Furthermore, EGFP-STIV could be used to evaluate viral replication conveniently under drug treatment, and the result showed that STIV replication was significantly inhibited after the addition of antioxidant pyrrolidine dithiocarbamate (PDTC). Thus, the EGFP-tagged recombinant iridovirus will not only be useful for further investigations on the viral replicative dynamics, but also provide an alternative simple strategy to screen for antiviral substances.
authors.affiliation: Key Laboratory of Marine Bio-resources Sustainable Utilization, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou 510301, PR China.
authors.forename: Youhua
authors.initials: Y
authors.lastname: Huang
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Huang
authors.forename: Jia
authors.initials: J
authors.lastname: Cai
authors.forename: Fuzhou
authors.initials: F
authors.lastname: Ye
authors.forename: Liya
authors.initials: L
authors.lastname: Guan
authors.forename: Hong
authors.initials: H
authors.lastname: Liu
authors.forename: Qiwei
authors.initials: Q
authors.lastname: Qin
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D004353:Drug Evaluation, Preclinical / Q000379:methods; D005818:Genetic Engineering; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D017994:Iridovirus / Q000235:genetics / Q000254:growth & development*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D013194:Staining and Labeling / Q000379:methods*; D014759:Viral Envelope Proteins / Q000235:genetics; D014773:Virology / Q000379:methods*; D014779:Virus Replication
chemical_list: D000998:Antiviral Agents; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins