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Construction of a reverse genetic system for porcine astrovirus.
Qin, Yifeng; Fang, Qingli; Liu, Huan; Ji, Chengyuan; Chen, Ying; Ouyang, Kang; Wei, Zuzhang; Huang, Weijian.
Afiliação
  • Qin Y; College of Animal Science and Technology, Guangxi University, No. 100 Daxue Road, Nanning, 530004, People's Republic of China.
  • Fang Q; College of Animal Science and Technology, Guangxi University, No. 100 Daxue Road, Nanning, 530004, People's Republic of China.
  • Liu H; College of Animal Science and Technology, Guangxi University, No. 100 Daxue Road, Nanning, 530004, People's Republic of China.
  • Ji C; College of Animal Science and Technology, Guangxi University, No. 100 Daxue Road, Nanning, 530004, People's Republic of China.
  • Chen Y; College of Animal Science and Technology, Guangxi University, No. 100 Daxue Road, Nanning, 530004, People's Republic of China.
  • Ouyang K; College of Animal Science and Technology, Guangxi University, No. 100 Daxue Road, Nanning, 530004, People's Republic of China.
  • Wei Z; College of Animal Science and Technology, Guangxi University, No. 100 Daxue Road, Nanning, 530004, People's Republic of China. zuzhangwei@gxu.edu.cn.
  • Huang W; College of Animal Science and Technology, Guangxi University, No. 100 Daxue Road, Nanning, 530004, People's Republic of China. huangweijian-1@163.com.
Arch Virol ; 163(6): 1511-1518, 2018 Jun.
Article em En | MEDLINE | ID: mdl-29450743
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.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Mamastrovirus / RNA Viral / Genoma Viral / Células Epiteliais / Genética Reversa Limite: Animals Idioma: En Revista: Arch Virol Ano de publicação: 2018 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Mamastrovirus / RNA Viral / Genoma Viral / Células Epiteliais / Genética Reversa Limite: Animals Idioma: En Revista: Arch Virol Ano de publicação: 2018 Tipo de documento: Article