RESUMO
Monitoring programs for highly dangerous avian diseases in the Russian Federation from 2001 to 2009 detected 77 samples that were PCR positive for avian paramyxovirus serotype-1 (APMV-1) from sick or dead feral and domestic pigeons. Nucleotide sequences of the fusion (F) gene, including a nucleotide sequence encoding the F protein cleavage site, were determined for these isolates. All of the studied isolates possessed virulent F0 protein cleavage sites (112KRKKRF117, 112RRQKRF117, or 112KRQKRF117). Intracerebral pathogenicity index (ICPI) values determined for seven of the isolates exceeded the value of 0.7 (the range from 0.8 to 1.41). Based on partial genome sequencing and phylogenetic analysis, the isolates were assigned to two individual sublineages within class II genotype VIb. It was determined that most of these Newcastle disease virus isolates (70/77) recovered from the pigeons belonged to a relatively poorly studied sublineage VIb/2. The complete nucleotide sequence of the genome for the Pigeon/Russia/Vladimir/687/05 isolate of sublineage VIb/2 was determined.
Assuntos
Columbidae , Genes Virais , Doença de Newcastle/virologia , Vírus da Doença de Newcastle/genética , Vírus da Doença de Newcastle/patogenicidade , Animais , Dados de Sequência Molecular , Vírus da Doença de Newcastle/classificação , Vírus da Doença de Newcastle/isolamento & purificação , Filogenia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Federação Russa , Análise de Sequência de Proteína/veterinária , Análise de Sequência de RNA/veterinária , Homologia de Sequência , VirulênciaRESUMO
Synthesis, cDNA cloning, and nucleotide sequencing of F gene of rinderpest virus strain K was carried out. Analysis of nucleotide sequence showed the only open reading frame coding for protein from 546 a.o. with mol. weight 58.6 kDa. The mean percentage of identical nucleotide residues between F genes of strains K, Kabete O, and L is 76.4% for 5'-untranslated region and 90.5% for translated region, the share of similar amino acid residues in the respective proteins is 92.9%. The structure of restriction site of F0 precursor protein in rinderpest strains with different virulence is similar. Protein F of rinderpest virus strain K has 3 potential glycosylation sites and 13 cystein residues in positions identical to those of F protein of rinderpest strains Kabete O and L.
Assuntos
Genes Virais , Glicoproteínas/genética , Vírus da Peste Bovina/genética , Proteínas Virais de Fusão/genética , Regiões 5' não Traduzidas , Sequência de Aminoácidos , Animais , Sequência de Bases , Bovinos , Clonagem Molecular , DNA Complementar , Glicoproteínas/química , Proteínas de Membrana , Dados de Sequência Molecular , Fases de Leitura Aberta , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido Nucleico , Proteínas Virais de Fusão/químicaRESUMO
Synthesis, cDNA cloning, and identification of H gene nucleotide sequence of rinderpest virus (RPV) K strain are carried out. Analysis of the identified nucleotide sequence has revealed the single open reading frame encoding a protein consisting of 609 amino acids with molecular weight of 68 kDa. The mean nucleotide homology between H genes of K, Kabete O and L strains in 88.0%, the mean amino acid homology of the corresponding proteins is 88.2%. RPV K strain hemagglutinin contains 5 potential glycosylation sites. The position of all 13 cystein bases is identical to positions in H proteins of RPV Kabete O and L strains. Studies of the hydrophobic profile of the compared proteins have shown 2 potential transmembrane fragments.
Assuntos
DNA Viral/genética , Vírus da Peste Bovina/genética , Animais , Sequência de Bases , Bovinos , Clonagem Molecular , DNA Complementar , Genes Virais , Dados de Sequência Molecular , Proteínas Estruturais Virais/genéticaRESUMO
The complete nucleotide sequence of HN gene, the region of F gene, and intergene regions (M-F, F-HN, and HN-L) of the BOR74 and BOR82 strains of Newcastle disease virus have been determined. Based on the nucleotide and amino acid sequences, the speeds of the nucleic and amino acid changes were calculated (approximately 10(-3) nucleotides or amino acids/year). The BOR strains were grouped phylogenetically with the asymptomatic strains. These strains and the BOR strains have the same motif of the cleavage site (112GKQGR116-L117), but the HN protein of BOR strains has the 572 amino acids which differ the BOR strains from all other strains (571, 577, and 616 amino acids).
Assuntos
Genes Virais , Proteína HN/genética , Vírus da Doença de Newcastle/genética , Proteínas Virais de Fusão/genética , Sequência de Bases , Genoma Viral , Dados de Sequência Molecular , FilogeniaRESUMO
A system for detection and strain differentiation of Newcastle disease virus (NDV) by reverse transcription of polymerase chain reaction (RT-PCR) (isolation of RNA, choice of primers for nested PCR, and purification of PCR products) and sequencing is developed and optimized. A nucleotide sequence of gene F site, coding for the F2/F1 cleavage site of F0 fusion protein and including several hypervariable regions, is determined for 10 Russian strains and vaccine strains. The data indicate a replacement of NDV populations in Russia and a rapid evolution of the virus. The origin of pathogenic NDV strains which have been circulating up to the present time is still unknown.
Assuntos
Vírus da Doença de Newcastle/classificação , Sequência de Bases , DNA Bacteriano , Dados de Sequência Molecular , Vírus da Doença de Newcastle/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Homologia de Sequência do Ácido Nucleico , Especificidade da EspécieRESUMO
A field isolate of Newcastle disease virus (NDV) was isolated in the Russko-Vysotskaya poultry farm, Leningrad region. Within four days after infection, the isolate caused 100% mortality in 60-day-old susceptible chickens. The HA titer of the allantoic fluid samples collected after one passage in SPF-chicken embryos was 1:512, and it reacted only with the NDV specific antiserum in HI test. Intracerebral pathogenicity index and mean embryo death time were 1.97 and 49 hours, respectively. The isolate has the amino acid sequence of the protease cleavage site of the fusion protein F0 (112R-R-Q-R-R-F117), which is similar to that in the velogenic strains of NDV. Therefore, it was concluded that the virus isolated in this work was an ethiological agent of the ND outbreak in this poultry farm.
Assuntos
Surtos de Doenças/veterinária , Doença de Newcastle/epidemiologia , Vírus da Doença de Newcastle/isolamento & purificação , Aves Domésticas/virologia , Animais , Domínio Catalítico/genética , Embrião de Galinha , Galinhas , Endopeptidases/metabolismo , Testes de Hemaglutinação , Doença de Newcastle/virologia , Vírus da Doença de Newcastle/genética , Vírus da Doença de Newcastle/metabolismo , Reação em Cadeia da Polimerase , RNA Viral/análise , Federação Russa/epidemiologia , Proteínas Virais de Fusão/genética , Proteínas Virais de Fusão/metabolismoRESUMO
Amplification of H-gene fragment in combination with cDNA nucleotide sequencing can be used for indication and strain differentiation of classical swine fever virus.