Complete nucleotide sequence of Middelburg virus, isolated from the spleen of a horse with severe clinical disease in Zimbabwe.
The complete nucleotide sequence of Middelburg virus (MIDV) was determined for strain MIDV-857 from Zimbabwe. The isolation of this virus in 1993 from a horse that died showing severe clinical signs represents the first indication that MIDV can cause severe disease in equids. Full-length cDNA copies of the viral genome were successfully synthesized by an innovative RT-PCR amplification approach using an 'anchor primer' combined with the SMART methodology described previously for the synthesis of full-length cDNA copies from genome segments of dsRNA viruses. The MIDV-857 genome is 11,674 nt, excluding the 5'-terminal cap structure and poly(A) tail (which varies in length from approximately 180 to approximately 220 residues). The organization of the genome is like that of other alphaviruses, including a read-through stop codon between the nsP3 and nsP4 genes. However, phylogenetic analyses of the structural protein amino acid sequences suggested that the MIDV E1 gene was generated by recombination with a Semliki Forest virus-like virus. This hypothesis was supported by bootscanning analysis using a recombination-detection program. The 3' untranslated region of MIDV-857 also contains a 112 nt duplication. This study reports the first full-length sequence of MIDV, which was obtained from a single RT-PCR product.
Attoui
Houssam
H
Department of Arbovirology, Institute for Animal Health, Pirbright, Woking, Surrey GU24 0NF, UK.
Sailleau
Corinne
C
Agence Française de Sécurité Sanitaire des Aliments, 22 rue Pierre Curie, 94703 Maisons-Alfort Cedex 07, France.
Mohd Jaafar
Fauziah
F
Department of Arbovirology, Institute for Animal Health, Pirbright, Woking, Surrey GU24 0NF, UK.
Belhouchet
Mourad
M
Unité de Virologie Moléculaire, Etablissement Français du Sang Alpes-Méditerranée, 149 Boulevard Baille, 13005 Marseille, France.
Biagini
Philippe
P
Unité de Virologie Moléculaire, Etablissement Français du Sang Alpes-Méditerranée, 149 Boulevard Baille, 13005 Marseille, France.
Cantaloube
Jean François
JF
Unité de Virologie Moléculaire, Etablissement Français du Sang Alpes-Méditerranée, 149 Boulevard Baille, 13005 Marseille, France.
de Micco
Philippe
P
Unité de Virologie Moléculaire, Etablissement Français du Sang Alpes-Méditerranée, 149 Boulevard Baille, 13005 Marseille, France.
Mertens
Peter
P
Department of Arbovirology, Institute for Animal Health, Pirbright, Woking, Surrey GU24 0NF, UK.
Zientara
Stephan
S
Agence Française de Sécurité Sanitaire des Aliments, 22 rue Pierre Curie, 94703 Maisons-Alfort Cedex 07, France.
D020413:3' Untranslated Regions / Q000235:genetics; D000524:Alphavirus / Q000235:genetics* / Q000302:isolation & purification; D018354:Alphavirus Infections / Q000662:veterinary; D000818:Animals; D001483:Base Sequence; D018388:Codon, Terminator / Q000235:genetics; D016679:Genome, Viral; D006734:Horse Diseases / Q000821:virology; D006736:Horses; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010802:Phylogeny; D012315:RNA Caps / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D011995:Recombination, Genetic; D012672:Semliki forest virus / Q000235:genetics; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013154:Spleen / Q000821:virology; D017361:Viral Nonstructural Proteins / Q000235:genetics; D015678:Viral Structural Proteins / Q000235:genetics; D015030:Zimbabwe
D020413:3' Untranslated Regions; D018388:Codon, Terminator; D012315:RNA Caps; D012333:RNA, Messenger; D017361:Viral Nonstructural Proteins; D015678:Viral Structural Proteins