Recombinant expression of Garlic virus C (GARV-C) capsid protein in insect cells and its potential for the production of specific antibodies

• Published in: Microbiological research Vol. 163; no. 3; pp. 354 - 361
• Main Authors: Alves-Júnior, Miguel, Menezes Marraccini, Fernanda, de Albuquerque Melo Filho, Péricles, Nepomuceno Dusi, André, Pio-Ribeiro, Gilvan, Morais Ribeiro, Bergmann
• Format: Journal Article
• Language: English
• Published: Germany Elsevier GmbH 01-01-2008
• Subjects: Allexivirus; Allium sativum; Allium spp; Animals; Antibodies, Viral - isolation & purification; Autographa californica; Blotting, Western; Brazil; Capsid protein; Capsid Proteins - biosynthesis; Capsid Proteins - chemistry; Capsid Proteins - genetics; Cell Line; Electrophoresis, Polyacrylamide Gel; Flexiviridae - genetics; Garlic - virology; Garlic virus c; GARV-C; Insecta; Molecular Weight; Nuclear polyhedrosis virus; Nucleopolyhedrovirus - genetics; Rabbits; Recombinant Proteins - biosynthesis; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Allium spp; Allexivirus; Capsid protein; GARV-C
• ISSN: 0944-5013; 1618-0623
• DOI: 10.1016/j.micres.2006.06.016

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 32
KDa 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.