About: A novel method to rescue and culture duck Astrovirus type 1 in vitro

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An Entity of Type : schema:ScholarlyArticle, within Data Space : covidontheweb.inria.fr associated with source document(s)

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	A novel method to rescue and culture duck Astrovirus type 1 in vitro
Creator	Chen, Junhao Jiang, Shijin Lan, Jingjing Li, Haie Lin, Shaoli Xie, Zhijing Yang, Yupeng Zhang, Ruihua
Source	PMC
abstract	BACKGROUND: Reverse genetics systems enable the manipulation of viral genomes and therefore serve as robust reverse genetic tools to study RNA viruses. A DNA-launched rescue system initiates the transcription of viral genomic cDNA from eukaryotic promoter in transfected cells, generating homogenous RNA transcripts in vitro and thus enhancing virus rescue efficiency. As one of the hazardous pathogens to ducklings, the current knowledge of the pathogenesis of duck astrovirus type 1 (DAstV-1) is limited. The construction of a DNA-launched rescue system can help to accelerate the study of the virus pathogenesis. However, there is no report of such a system for DAstV-1. METHODS: In this study, a DNA-launched infectious clone of DAstV-1 was constructed from a cDNA plasmid, which contains a viral cDNA sequence flanked by hammerhead ribozyme (HamRz) and a hepatitis delta virus ribozyme (HdvRz) sequence at both terminals of the viral genome. A silent nucleotide mutation creating a Bgl II site in the ORF2 gene was made to distinguish the rescued virus (rDAstV-1) from the parental virus (pDAstV-1). Immunofluorescence assay (IFA) and western blot were conducted for rescued virus identification in duck embryo fibroblast (DEF) cells pre-treated with trypsin. The growth characteristics of rDAstV-1 and pDAstV-1 in DEF cells and the tissue tropism in 2-day-old ducklings of rDAstV-1 and pDAstV-1 were determined. RESULTS: The infectious DAstV-1 was successfully rescued from baby hamster kidney (BHK-21) cells and could propagate in DEF cells pre-treated with 1 μg/ml trypsin. Upon infection of DEF cells pre-treated with trypsin, DAstV-1 mRNA copies were identified after serial passaging, and the result showed that rDAstV-1 and pDAstV-1 shared similar replication kinetics. Animal experiment showed that the rDAstV-1 had an extensive tissue tropism, and the virus was capable of invading both the central and the peripheral immune organs in infected ducklings. CONCLUSIONS: An improved DNA-launched reverse genetics system for DAstV-1 was firstly constructed. Infectious virus recovered from BHK-21 cells could propagate in DEF cells pre-treated with trypsin. This is the first report of the successful in vitro cultivation of DAstV-1. We believe this valuable experimental system will contribute to the further study of DAstV-1 genome function and pathogenesis.
has issue date	2019-09-05(xsd:dateTime)
bibo:doi	10.1186/s12985-019-1218-5
bibo:pmid	31488178
has license	cc-by
sha1sum (hex)	ab49a01b79f3623438ca5e7f5833d95ef1701a55
schema:url	https://doi.org/10.1186/s12985-019-1218-5
resource representing a document's title	A novel method to rescue and culture duck Astrovirus type 1 in vitro
has PubMed Central identifier	PMC6729042
has PubMed identifier	31488178
schema:publication	Virol J
resource representing a document's body	covid:ab49a01b79f3623438ca5e7f5833d95ef1701a55#body_text
is schema:about of	named entity 'cells' named entity 'central' named entity 'ducklings' named entity 'trypsin' named entity 'replication' named entity 'Immunofluorescence' named entity 'robust' named entity 'rescued' named entity 'embryo' named entity 'FLANKED' named entity 'VIRUS IDENTIFICATION' named entity 'INFECTION' named entity 'GENE' named entity 'METHOD' named entity 'PATHOGENS' named entity 'METHODS' named entity 'PRE' named entity 'ACCELERATE' named entity 'ORGANS' named entity 'INFECTIOUS' named entity 'RESULT' named entity 'WESTERN BLOT' named entity 'IN VITRO' named entity 'RNA' named entity 'EMBRYO FIBROBLAST' named entity 'TRANSCRIPTS' named entity 'CDNA' named entity 'BABY' named entity 'RNA VIRUSES' named entity 'RESCUE' named entity 'A HEPATITIS' named entity 'CAPABLE' named entity 'SITE' named entity 'SILENT' named entity 'STUDY' named entity 'IMMUNOFLUORESCENCE ASSAY' named entity 'A DNA' named entity 'TOOLS' named entity 'GENERATING' named entity 'EUKARYOTIC PROMOTER' named entity 'MANIPULATION' named entity 'DUCK EMBRYO' named entity 'GENOMIC' named entity 'RIBOZYME' named entity 'TYPE 1' named entity 'TYPE 1' named entity 'ROBUST' named entity 'LIMITED' named entity 'HAMMERHEAD RIBOZYME' named entity 'EXTENSIVE' named entity 'HELP' named entity 'TRANSCRIPTION' named entity 'SHARED' named entity 'RESCUE' named entity 'CULTURE' named entity 'NOVEL' named entity 'SERIAL' named entity 'TRYPSIN' named entity 'IMMUNE' named entity 'GENETIC' named entity 'CONSTRUCTION' named entity 'BACKGROUND' named entity 'REVERSE GENETICS' named entity 'NUCLEOTIDE MUTATION' named entity 'HAMSTER' named entity 'COPIES' named entity 'REPLICATION' named entity 'GROWTH CHARACTERISTICS' named entity 'REVERSE' named entity 'SIMILAR' named entity 'DEF' named entity 'ONE OF' named entity 'BCG' named entity 'ENABLE'

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