A first full outer capsid protein sequence data-set in the Orbivirus genus (family Reoviridae): cloning, sequencing, expression and analysis of a complete set of full-length outer capsid VP2 genes of the nine African horsesickness virus serotypes.
Abstract: The outer capsid protein VP2 of African horsesickness virus (AHSV) is a major protective antigen. We have cloned full-length VP2 genes from the reference strains of each of the nine AHSV serotypes. Baculovirus recombinants expressing the cloned VP2 genes of serotypes 1, 2, 4, 6, 7 and 8 were constructed, confirming that they all have full open reading frames. This work completes the cloning and expression of the first full set of AHSV VP2 genes. The clones of VP2 genes of serotypes 1, 2, 5, 7 and 8 were sequenced and their amino acid sequences were deduced. Our sequencing data, together with that of the published VP2 genes of serotypes 3, 4, 6 and 9, were used to generate the first complete sequence analysis of all the (sero)types for a species of the Orbivirus genus. Multiple alignment of the VP2 protein sequences showed that homology between all nine AHSV serotypes varied between 47.6 % and 71.4 %, indicating that VP2 is the most variable AHSV protein. Phylogenetic analysis grouped together the AHSV VP2s of serotypes that cross-react serologically. Low identity between serotypes was demonstrated for specific regions within the VP2 amino acid sequences that have been shown to be antigenic and play a role in virus neutralization. The data presented here impact on the development of new vaccines, the identification and characterization of antigenic regions, the development of more rapid molecular methods for serotype identification and the generation of comprehensive databases to support the diagnosis, epidemiology and surveillance of AHS.
Publication Date: 2003-04-15 PubMed ID: 12692299DOI: 10.1099/vir.0.18919-0Google Scholar: Lookup
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Summary
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The research paper focuses on the cloning, sequencing, expression and analysis of outer capsid VP2 genes of African horsesickness virus (AHSV). This represents the first complete sequence analysis for every distinct type (or serotype) of a species under the Orbivirus genus. The data could influence the development of new vaccines and rapid methods for identifying different serotypes.
Gene Cloning, Sequencing and Expression
- The study successfully cloned full-length VP2 genes from the reference strains of all nine AHSV serotypes.
- Baculovirus recombinants were used to express the cloned VP2 genes of serotypes 1, 2, 4, 6, 7 and 8, confirming these serotypes all have full open reading frames. This completes the cloning and expression of the first full set of AHSV VP2 genes.
Sequence Analysis and Comparison
- The team performed sequence analysis of the cloned VP2 genes. This combined with existing data on VP2 genes of serotypes 3, 4, 6 and 9 allowed the generation of the first complete sequence analysis for a species of the Orbivirus genus.
- Multiple alignment of the VP2 protein sequences revealed that homology between all nine AHSV serotypes ranged from 47.6% to 71.4%, suggesting that VP2 is the most variable protein in AHSV.
Phylogenetic Analysis and Implications to Vaccine Development
- Phylogenetic analysis showed that AHSV VP2s of serotypes grouping together based on serological cross-reactivity.
- Specific regions within VP2 amino acid sequences have been identified, exhibiting low identity between serotypes. These regions are antigenic and play a crucial role in virus neutralization. Therefore, the knowledge obtained from this study is expected to facilitate vaccine development and the creation of rapid molecular methods for serotype identification.
- This information is crucial for the development of a comprehensive database that will help in diagnosing, understanding the epidemiology and effectively monitoring AHS.
Cite This Article
APA
Potgieter AC, Cloete M, Pretorius PJ, van Dijk AA.
(2003).
A first full outer capsid protein sequence data-set in the Orbivirus genus (family Reoviridae): cloning, sequencing, expression and analysis of a complete set of full-length outer capsid VP2 genes of the nine African horsesickness virus serotypes.
J Gen Virol, 84(Pt 5), 1317-1326.
https://doi.org/10.1099/vir.0.18919-0 Publication
Researcher Affiliations
- Onderstepoort Veterinary Institute, 0110 Onderstepoort, South Africa.
- Onderstepoort Veterinary Institute, 0110 Onderstepoort, South Africa.
- Division of Biochemistry, School for Chemistry and Biochemistry, University of Potchefstroom for CHE, 2520 Potchefstroom, South Africa.
- Onderstepoort Veterinary Institute, 0110 Onderstepoort, South Africa.
MeSH Terms
- African Horse Sickness / virology
- African Horse Sickness Virus / classification
- African Horse Sickness Virus / genetics
- African Horse Sickness Virus / metabolism
- Amino Acid Sequence
- Animals
- Baculoviridae / genetics
- Baculoviridae / metabolism
- Capsid Proteins / chemistry
- Capsid Proteins / genetics
- Capsid Proteins / metabolism
- Cloning, Molecular
- Horses
- Mice
- Molecular Sequence Data
- Orbivirus / classification
- Orbivirus / genetics
- Phylogeny
- Recombination, Genetic
- Sequence Alignment
- Sequence Analysis, DNA
- Serotyping
Citations
This article has been cited 5 times.- van Gennip RGP, van de Water SGP, Potgieter CA, van Rijn PA. Structural Protein VP2 of African Horse Sickness Virus Is Not Essential for Virus Replication In Vitro.. J Virol 2017 Feb 15;91(4).
- Potgieter AC, Wright IM, van Dijk AA. Consensus Sequence of 27 African Horse Sickness Virus Genomes from Viruses Collected over a 76-Year Period (1933 to 2009).. Genome Announc 2015 Sep 10;3(5).
- Kanai Y, van Rijn PA, Maris-Veldhuis M, Kaname Y, Athmaram TN, Roy P. Immunogenicity of recombinant VP2 proteins of all nine serotypes of African horse sickness virus.. Vaccine 2014 Sep 3;32(39):4932-7.
- Bachanek-Bankowska K, Maan S, Castillo-Olivares J, Manning NM, Maan NS, Potgieter AC, Di Nardo A, Sutton G, Batten C, Mertens PP. Real time RT-PCR assays for detection and typing of African horse sickness virus.. PLoS One 2014;9(4):e93758.
- Maan NS, Maan S, Nomikou K, Belaganahalli MN, Bachanek-Bankowska K, Mertens PP. Serotype specific primers and gel-based RT-PCR assays for 'typing' African horse sickness virus: identification of strains from Africa.. PLoS One 2011;6(10):e25686.
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