期刊
VIROLOGY
卷 325, 期 2, 页码 308-319出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.virol.2004.04.046
关键词
PRRSV; nucleotide sequence; full-length cDNA; in vitro transcripts; transfection; MARC-145 cells; primer extension; PRRSV pathogenesis
类别
资金
- NCRR NIH HHS [P20 RR015635, P20RR15635] Funding Source: Medline
- NIAID NIH HHS [AI-34956, R01 AI034956, R29 AI034956] Funding Source: Medline
The nucleotide sequence of a highly pathogenic porcine reproductive and respiratory syndrome virus (PRRSV) was determined. Transfection of MARC-145 cells with capped in vitro transcripts derived from a full-length cDNA clone of the viral genome resulted in infectious PRRSV with growth characteristics similar to that of the parental virus. Primer extension analysis revealed that during replication, the viral polymerase corrected the two nonviral guanosine residues present at the 5' terminus of the transfected transcripts. Animal studies showed that the cloned virus induced hyperthermia, persistent viremia, and antibody response, similar to that observed with the parental virus. Contact transmission occurred rapidly within 3 days of introduction of naive pigs into the group of clone virus-inoculated pigs. These results suggest that the cloned virus retains the in vivo virulence and contagion properties of the parental virus, thus, providing the background for reverse genetics manipulation in systematic examination of attenuation and virulence phenotypes. (C) 2004 Elsevier Inc. All rights reserved.
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