Recombinant expression of Garlic virus C (GARV-C) capsid protein in insect cells and its potential for the production of specific antibodies.
Garlic cultivars in Brazil are infected by a complex of viruses and for some virus species, such as the allexivirus, purification of the virions is sometimes cumbersume. To overcome this problem, recombinant expression of viral proteins in heterologous systems is an alternative method for producing antibodies. The capsid gene from Garlic virus C (GarV-C), an Allexivirus, was inserted into the genome of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) generating the recombinant virus vSynGarV-C. The recombinant protein expression was confirmed by SDS-PAGE and western-blot of extracts from recombinant virus infected insect cells, where a protein band of approximately 32KDa was observed only in extracts from recombinant infected cells. This protein corresponded to the predicted size of the capsid protein of the GarV-C. A rabbit polyclonal antibody was raised against this protein, shown to be specific for the GarV-C protein in western-blot and dot-Elisa, however with a low titer.
Alves-Júnior
Miguel
M
Department of Agronomy, Universidade Federal Rural de Pernambuco, Recife, PE, Brazil.
Menezes Marraccini
Fernanda
F
Melo Filho
Péricles de Albuquerque
Pde A
Nepomuceno Dusi
André
A
Pio-Ribeiro
Gilvan
G
Morais Ribeiro
Bergmann
B
D000818:Animals; D000914:Antibodies, Viral / Q000302:isolation & purification; D015153:Blotting, Western; D001938:Brazil; D036022:Capsid Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D052677:Flexiviridae / Q000235:genetics*; D005737:Garlic / Q000821:virology; D007313:Insecta; D008970:Molecular Weight; D017924:Nucleopolyhedroviruses / Q000235:genetics; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics
D000914:Antibodies, Viral; D036022:Capsid Proteins; D011994:Recombinant Proteins