RESUMO
A 0.9 kb cDNA for the foot and mouth disease virus (FMDV) type Asia 1 63/72, cloned in the plasmid pUR222 by dC/dG tailing method, was expressed into a protein which was immunogenic in guinea pigs and cattle. The protein purified to homogeneity was found to be basic and of 38 kDa. A sequence of 879 nucleotides of the inserted cDNA was obtained. The nucleotide sequence was 65% GC-rich and was homologous to the gene for VPI of FMDV types A5, OIK and C3 to the extent of 35-40%. From the nucleotide sequence, a sequence of 293 amino acids was derived which contained 43 arginine, 4 lysine, 7 glutamic acid and 18 aspartic acid residues making the protein highly basic. The molecular weight was calculated to be 31.6 kDa. The 38 kDa protein produced by the cloned cDNA is a fused protein composed of the 293 amino acids; 5 and 55 amino acids of the alpha-complementation protein of the beta-galactosidase at the N and C terminal, respectively, and 5 amino acid coded by the dG/dC tails used for cloning the cDNA.
Assuntos
Sequência de Aminoácidos , Antígenos Virais/genética , Aphthovirus/química , Sequência de Bases , DNA Viral/isolamento & purificação , Dados de Sequência Molecular , Proteínas Estruturais Virais/genéticaRESUMO
A clone bearing a 9.8 kb EcoRI fragment of rice DNA containing the genes for the rRNAs and the intergenic spacer was identified by screening a rice genomic library in lambda Charon 4 phage with rRNAs. The 9.8 kb EcoRIDNA fragment was found to be a minor rDNA unit of rice variety IR-20. The rRNA genes and the intergenic spacer were mapped by hybridization and nucleotide sequence analyses. The DNAs in the intergenic spacer of the minor rDNA unit of 9.8 kb and the major rDNA unit of 8.9 kb cross-hybridized showing that those regions are homologous.