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Archives of Virology
Published in: Archives of Virology 6/2018

01-06-2018 | Original Article

Construction of a reverse genetic system for porcine astrovirus

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

Published in: Archives of Virology | Issue 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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Metadata
Title
Construction of a reverse genetic system for porcine astrovirus
Authors
Yifeng Qin
Qingli Fang
Huan Liu
Chengyuan Ji
Ying Chen
Kang Ouyang
Zuzhang Wei
Weijian Huang
Publication date
01-06-2018
Publisher
Springer Vienna
Published in
Archives of Virology / Issue 6/2018
Print ISSN: 0304-8608
Electronic ISSN: 1432-8798
DOI
https://doi.org/10.1007/s00705-018-3771-4

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