Production of recombinant Potato mop-top virus coat protein in Escherichia coli and generation of antisera recognising native virus protein.
Potato mop-top virus (PMTV, Pomovirus) is difficult to detect because it is unevenly distributed and present at low concentration in infected tissues. The production of PMTV-free seed relies on sensitive and specific detection methods of virus detection, including serological methods. The possibility of using a PMTV recombinant coat protein (CP) as an antigen for antiserum production was investigated. The region encoding the PMTV CP was inserted into pET3A, expressed in Escherichia coli, and the recombinant PMTV CP produced was used to raise antibodies in rabbits. Three antisera were produced. All recognised efficiently the recombinant CP in Western blot analysis and the most sensitive antiserum (H5003) detected native CP on Western blots and in ELISA. Thus, recombinant CP can be used as an alternative to purified virus for the production of specific antibodies against PMTV.
Hélias
Valérie
V
FNPPPT, Fédération Nationale des Producteurs de Plants de Pommes de Terre, 9 rue d'Athènes, 75009, Paris, France.
Jacquot
Emmanuel
E
Guillet
Maryse
M
Le Hingrat
Yves
Y
Giblot-Ducray
Danièle
D
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis / Q000276:immunology*; D015153:Blotting, Western; D036022:Capsid Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D007114:Immunization; D010935:Plant Diseases / Q000821:virology; D010942:Plant Viruses / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D012328:RNA Viruses / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D012680:Sensitivity and Specificity; D011198:Solanum tuberosum / Q000821:virology*
D000914:Antibodies, Viral; D036022:Capsid Proteins; D011994:Recombinant Proteins