Molecular cloning, sequencing and expression in Escherichia coli of capsid protein gene from rabbit haemorrhagic disease virus (Spanish isolate AST/89)

Boga, J.A.; Casais, R.; Marin, M.S.; Martin Alonso, J.M.; Carmenes, R.S.; Prieto, M.; Parra, F.

Journal of General Virology 75(9): 2409-2413


ISSN/ISBN: 0022-1317
Accession: 002436192

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The major capsid component (VP60) from the Spanish field isolate AST/89 of rabbit haemorrhagic disease virus (RHDV) was cloned, sequenced and expressed in E. coli. The sequence of the 3'-terminal 2483 nucleotides of the genome was found to be 95.4% identical to the German RHDV strain, showing 10 changes in the deduced VP60 amino acid sequence. The gene coding for this structural polypeptide was expressed in bacteria as a beta -galactosidase fusion protein or using a T7 RNA polymerase-based system. The VP60 fusion protein showed only partial antigenic similarity with native VP60 and did not confer protective immunity. The recombinant VP60 produced in the T7 RNA polymerase-based system was antigenically similar to the viral polypeptide as determined using polyclonal and monoclonal antibodies. When used to immunize rabbits the recombinant VP60 was able to protect the animals against a lethal challenge using purified RHDV.