Molecular cloning, sequencing and expression in Escherichia coli of the capsid protein gene from rabbit haemorrhagic disease virus (Spanish isolate AST/89)
| Autor: | Miguel A. Prieto, José Antonio Boga, José M. Martín-Alonso, M. S. Marin, Rosa Casais, R. S. Carmenes, Francisco Parra |
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| Rok vydání: | 1994 |
| Předmět: |
Genes
Viral Hemorrhagic Disease Virus Rabbit Recombinant Fusion Proteins Genetic Vectors Molecular Sequence Data Restriction Mapping Gene Expression Molecular cloning Polymerase Chain Reaction Virus law.invention Rabbit haemorrhagic disease Capsid law Germany Virology Escherichia coli medicine Animals T7 RNA polymerase Cloning Molecular Gene DNA Primers Viral Structural Proteins Base Sequence biology biology.organism_classification Fusion protein Molecular biology Spain Recombinant DNA Electrophoresis Polyacrylamide Gel Rabbits medicine.drug |
| Zdroj: | Scopus-Elsevier |
| ISSN: | 1465-2099 0022-1317 |
| DOI: | 10.1099/0022-1317-75-9-2409 |
| Popis: | We describe the cloning, nucleotide sequencing and expression in Escherichia coli of the major capsid component (VP60) from the Spanish field isolate AST/89 of rabbit haemorrhagic disease virus (RHDV). The sequence of the 3'-terminal 2483 nucleotides of the genome was found to be 95.4% identical to the German RHDV strain, showing ten changes in the deduced VP60 amino acid sequence. The gene coding for this structural polypeptide has been 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. |
| Databáze: | OpenAIRE |
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