We found a match
Your institution may have rights to this item. Sign in to continue.
- Title
Construction of a reverse genetic system for porcine astrovirus.
- Authors
Qin, Yifeng; Fang, Qingli; Liu, Huan; Ji, Chengyuan; Chen, Ying; Ouyang, Kang; Wei, Zuzhang; Huang, Weijian
- 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.
- Subjects
ASTROVIRUSES; ANTISENSE DNA; POLYMERASE chain reaction; PATHOGENIC microorganisms; GENE transfection
- Publication
Archives of Virology, 2018, Vol 163, Issue 6, p1511
- ISSN
0304-8608
- Publication type
Article
- DOI
10.1007/s00705-018-3771-4