In vitro expression and monoclonal antibody of RNA-dependent RNA polymerase for infectious bursal disease virus.
VP1, the RNA-dependent RNA polymerase of infectious bursal disease virus (IBDV), has been suggested to play an essential role in the replication and translation of viral RNAs. In this study, we first expressed the complete VP1 protein gene in Escherichia coli (E. coli), and then the produced polyclonal antibody and four monoclonal antibodies (mAbs) to recombinant VP1 protein (rVP1) were shown to bind the IBDV particles in chicken embryo fibroblast and Vero cells. The epitopic analysis showed that mAbs 1D4 and 3C7 recognized respectively two distinct antigenic epitopes on the rVP1 protein, but two pair of mAbs 1A2/2A12 and 1E1/1H3 potentially recognized another two topologically related epitopes. Immunocytochemical stainings showed that VP1 protein formed irregularly shaped particles in the cytoplasm of the IBDV-infected cells. These results demonstrated that the mAbs to rVP1 protein could bind the epitopes of IBDV particles, indicating that the rVP1 protein expressed in E. coli was suitable for producing the mAb to VP1 protein of IBDV, and that the cytoplasm could be the crucial site for viral genome replication of IBDV.
Zheng
Xiaojuan
X
Laboratory of Virology and Immunology, Institute of Preventive Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou, Zhejiang, People's Republic of China.
Hong
Lianlian
L
Li
Yifei
Y
Guo
Junqing
J
Zhang
Gaiping
G
Zhou
Jiyong
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology; D002642:Chick Embryo; D002522:Chlorocebus aethiops; D000939:Epitopes; D007243:Infectious bursal disease virus / Q000201:enzymology* / Q000235:genetics; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D014162:Transfection; D014709:Vero Cells; D015678:Viral Structural Proteins / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000939:Epitopes; D011993:Recombinant Fusion Proteins; C107669:VP1 protein, infectious bursal disease virus; D015678:Viral Structural Proteins; D012324:RNA-Dependent RNA Polymerase