Inhibition of potato virus Y NIa activity: preparation of monoclonal antibody directed against PVY NI protein that inhibits cleavage of PVY polyprotein.
A partially purified nuclear inclusion (NI) fraction was obtained from tobacco plants infected by potato virus Y (PVY). Four monoclonal antibodies (MAbs) were produced and characterized using this semipurified fraction as antigen. Data showed that only one was directed against NIa whereas two were directed against cytoplasmic inclusion (CI) protein and the last one against coat protein (CP). These results were due to the fact that the semipurified NI fraction was usually contaminated with CI and CP proteins. When used on in situ immunofluorescence method the anti-NIa MAb showed accumulation of the NIa protein in both nucleus and cytoplasm. In vivo, this MAb was able to detect different forms of the NIa protein including precursors and cleavage products. It was also able to inhibit the cleavage of the polyprotein detected in the semipurified NI.
Rouis
S
S
Centre de Biotechnologie de Sfax, Tunisia.
Traincard
F
F
Gargouri
R
R
Dartevelle
S
S
Jeannequin
O
O
Mazié
J C
JC
Ayadi
H
H
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D015153:Blotting, Western; D002213:Capsid / Q000276:immunology; D010450:Endopeptidases / Q000032:analysis / Q000096:biosynthesis / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D051379:Mice; D010936:Plant Extracts / Q000737:chemistry; D010947:Plants, Toxic; D017800:Potyvirus / Q000201:enzymology / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000096:biosynthesis; D014026:Nicotiana / Q000821:virology*; D014764:Viral Proteins / Q000032:analysis / Q000096:biosynthesis / Q000276:immunology*
D000911:Antibodies, Monoclonal; D010936:Plant Extracts; D011994:Recombinant Proteins; D014764:Viral Proteins; C406275:cylindrical inclusion protein, potyvirus; D010450:Endopeptidases; C059992:nuclear inclusion protein a, mosaic viruses