RESUMO
Mammalian 2'-5' oligoadenylate (2-5A) synthetases are important mediators of the antiviral activity of interferons. Both human and mouse 2-5A synthetase gene families encode four forms of enzymes: small, medium, large and ubiquitin-like. In this study, the structures of four equine OAS genes were determined using DNA sequences derived from fifteen cDNA and four BAC clones. Composition of the equine OAS gene family is more similar to that of the human OAS family than the mouse Oas family. Two OAS-containing bovine BAC clones were identified in GenBank. Both equine and bovine BAC clones were physically assigned by FISH to horse and cattle chromosomes, ECA8p15-->p14 and BTA17q24--> q25, respectively. The comparative mapping data confirm conservation of synteny between ungulates, humans and rodents.
Assuntos
2',5'-Oligoadenilato Sintetase/genética , Cavalos/genética , Animais , Mapeamento Cromossômico , Cromossomos Artificiais Bacterianos , Primers do DNA , DNA Complementar/genética , Etiquetas de Sequências Expressas , Hibridização in Situ Fluorescente , Família Multigênica , RNA de Cadeia Dupla/genéticaRESUMO
West Nile (WN) virus is the most widespread among flaviviruses, but until recently it was not known on the American continent. We describe here design of a subgenomic replicon, as well as a full-length infectious clone of the lineage II WN strain, which appeared surprisingly stable compared to other flavivirus infectious clones. This infectious clone was used to investigate effects of 5'- and 3'-nonrelated sequences on virus replication and infectivity of synthetic RNA. While a long nonrelated sequence at the 3'-end delayed but did not prevent establishment of the productive infectious cycle, a much shorter extra sequence at the 5'-end completely abrogated virus replication. Replacement of the conserved 5'-adenosine residue substantially delayed, but did not prevent, establishment of virus infection. In all cases, the recovered virus had restored its authentic 5'- and 3'-end genome sequences. However, the presence of extensive nonrelated sequences at both 5'- and 3'-ends could not be repaired.
Assuntos
Genoma Viral , Vírus do Nilo Ocidental/genética , Regiões 3' não Traduzidas/genética , Regiões 5' não Traduzidas/genética , Linhagem Celular , Clonagem Molecular , Escherichia coli/genética , Replicon , Transcrição Gênica , Transfecção , Replicação Viral/genética , Vírus do Nilo Ocidental/patogenicidadeRESUMO
A 323-bp DNA fragment (U15557) was isolated, cloned, and sequenced after polymerase chain reaction (PCR) amplification from Monodelphis domestica genomic DNA. A HindIII restriction fragment length polymorphism was identified in this species using the U15557 PCR, fragment as a hybridization probe. DNA samples exhibited either a 6.4 kb band, a 7.2 kb band, or both bands simultaneously. Behaviour of these two variants in family studies was consistent with codominant autosomal inheritance. Linkage between this marker and the loci encoding protease inhibitor (PI) and adenylate kinase 1 (AK1) was found in M. domestica.
Assuntos
Adenilato Quinase/genética , Ligação Genética , Marcadores Genéticos/genética , Gambás/genética , Inibidores de Proteases , Animais , Clonagem Molecular , Cruzamentos Genéticos , Feminino , Masculino , Dados de Sequência Molecular , Polimorfismo de Fragmento de Restrição , Análise de Sequência de DNA , Fatores SexuaisRESUMO
A panel of 25 microsatellite loci known to map to human chromosome 18 was screened for length polymorphisms in a multigenerational pedigree of baboons (Papio hamadryas). Eight of these loci were polymorphic in the baboon pedigrees, with observed heterozygosity values ranging from 0.16 to 0.88. All eight loci show strong evidence of linkage. The most likely map order among the microsatellite loci for Papio is completely compatible with the locus order found in the human genome, and sex differences in recombination rates are also similar in the two species. We conclude that the organization of the chromosomal region defined by these loci is largely conserved between baboons and humans.
