Cytopathic effect of classical swine fever virus NS3 protein on PK-15 cells.
In order to further research the relationship between classical swine fever virus' (CSFV) NS3 protein and the cytopathic effect (CPE) in cells infected with the CSFV, and to reveal the effect of protein NS3 on the host cells, the NS3 of CSFV Shimen strain amplified by RT-PCR was subcloned into the pEGFP-C1, named pEGFP-C1-NS3. The insert position, the size and the reading frame were correct for restriction enzyme digestion and sequence analysis. The pEGFP-C1-NS3 and pEGFP-C1 were transfected into PK-15 cells by liposome, and positive cell clones were gained by G418. The NS3-EGFP fusion protein expressed in pEGFP-C1-NS3 cells was observed by inverted fluorescence microscopy and identified by Western blot. The CPE appeared in positive pEGFP-C1-NS3 cells 72 h after passaging, apoptosis detection was also performed on positive pEGFP-C1-NS3 cells and pEGFP-C1 cells 72 h after passaging by TUNEL assay. The apoptosis rates in the positive pEGFP-C1-NS3 and pEGFP-C1 cells were 43.4 and 13.1%, respectively (p < 0.05). The results suggest that the CPE in positive pEGFP-C1-NS3 cells was induced by apoptosis and there is a relationship between the expression of NS3 and apoptosis.
Xu
Hao
H
College of Veterinary Medicine, Northwest A&F University, Yangling, China.
Hong
Hai-Xia
HX
Zhang
Yan-Ming
YM
Guo
Kang-Kang
KK
Deng
Xiao-Ming
XM
Ye
Gui-Sheng
GS
Yang
Xiao-Yun
XY
D000818:Animals; D017209:Apoptosis / Q000502:physiology; D002478:Cells, Cultured / Q000187:drug effects* / Q000473:pathology; D006691:Classical Swine Fever / Q000821:virology; D006692:Classical Swine Fever Virus / Q000737:chemistry*; D003588:Cytopathogenic Effect, Viral; D010957:Plasmids / Q000235:genetics; D013552:Swine; D017361:Viral Nonstructural Proteins / Q000494:pharmacology*
D017361:Viral Nonstructural Proteins