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Erschienen in: Archives of Virology 6/2018

15.02.2018 | Original Article

Construction of a reverse genetic system for porcine astrovirus

verfasst von: Yifeng Qin, Qingli Fang, Huan Liu, Chengyuan Ji, Ying Chen, Kang Ouyang, Zuzhang Wei, Weijian Huang

Erschienen in: Archives of Virology | Ausgabe 6/2018

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Abstract

In order to construct a full-length infectious cDNA clone of porcine astrovirus, three fragments covering the complete genome of PAstV1-GX1 strain were amplified by RT-PCR. All three PCR-amplified fragments were cloned into T-Vector pMD19 (Simple), and subsequently assembled into a full-length cDNA clone by subcloning. A silent nucleotide change creating a PstI site was engineered into the full-length cDNA clone to distinguish the rescued virus from the parental virus. Upon transfection of BHK-21 cells with the in vitro transcripts of both the original and constructed cDNAs, typical cytopathic effects were observed on PK-15 cells after serial passaging of the cell supernatant. The construction and recovery of the infectious cDNA clone of porcine astrovirus will provide a valuable experimental system to study the genome function and pathogenesis of astroviruses.
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Metadaten
Titel
Construction of a reverse genetic system for porcine astrovirus
verfasst von
Yifeng Qin
Qingli Fang
Huan Liu
Chengyuan Ji
Ying Chen
Kang Ouyang
Zuzhang Wei
Weijian Huang
Publikationsdatum
15.02.2018
Verlag
Springer Vienna
Erschienen in
Archives of Virology / Ausgabe 6/2018
Print ISSN: 0304-8608
Elektronische ISSN: 1432-8798
DOI
https://doi.org/10.1007/s00705-018-3771-4

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