Construction of cytopathic PK-15 cell model of classical swine fever virus
No cytopathic effect (CPE) can be observed on classical swine fever virus (CSFV) infected cell culture in vitro. This brings an obstacle to the researches on reciprocity between CSFV and host cells. Based on the construction of full-length genomic infectious cDNA clone of Chinese CSFV standard virulent Shimen strain, partial deletion is introduced into genomic cDNA to obtain a 7.5 kb subgenomic cDNA. A new subgenomic CSFV is derived from transfection with the subgenomic cDNA on PK-15 cells pre-infected by CSFV Shimen virus. Typical CPE induced by this subgenomic virus is observed on PK-15 cells. Coexistence of wildtype and subgenomic virus in cytopathic cell culture is demonstrated by RT-PCR detection in cytopathic cells. For conclusion, the construction of cytopathic cell model exploited a new way for researches on the molecular mechanism of CSFV pathogenesis.
Keywordsclassical swine fever virus genomic infectious clone subgenome defective interfering particle cytopathic effect
Unable to display preview. Download preview PDF.
- 1.Yin, Z, Liu, J, Animal Virology (in Chinese), 2nd ed., Beijing: Science Press, 1997, 652–664.Google Scholar
- 2.Wang, Z. S, Look back and prospect of prevention and control of classical swine fever, in Proceeding of China Institute of Veterinary Drug Control (eds. Chen, Y, Zheng, M, Wang, Z.} in Chinese), Beijing: China Institute of Veterinary Drug Control, 1999, 1–28.Google Scholar
- 4.Christian, M, Peter, S, Jon-duri, T. et al., Cytopathogenic and noncytopathogenic RNA replicons of classical swine fever virus, J. Virol., 1999, 73: 7787–7794.Google Scholar
- 5.Meyers, G, Thiel, H. J., Cytopathogenicity of classical swine fever virus caused by defective interfering particles, J. Virol., 1995, 69: 3683–3689.Google Scholar
- 8.Vassilev, V. B, Collett, M. S, Donis, R. O., Authentic and chimeric full-length genomic cDNA clones of bovine viral diarrhea virus that yield infectious transcripts, J. Virol., 1997, 71: 471–478.Google Scholar
- 10.Moorann, R. J. M, Van Gennip, H. G. P, Miedema, G. K. W. et al., Infectious RNA transcribed from an engineered full-length cDNA template of the genome of a pestivirus, J. Virol., 1996, 70: 763–770.Google Scholar
- 11.Ruggli, N, Tratschin, J. D, Mittelholzer, C. et al., Nucleotide sequence of classical swine fever virus strain Alfort/187 and transcription of infectious RNA from stably cloned full-length cDNA, J. Virol., 1996, 70: 3478–3487.Google Scholar
- 14.Fang, B, Zhao, W, Hu, Z., Long template PCR amplification of 5′ half molecule of Chinese type 2 Dengue virus, Periodicals of Academy of Military Medical Science (in Chinese), 2000, 24: 274.Google Scholar