RESUMO
The epidemiologically important liver flukes Opisthorchis felineus, Opisthorchis viverrini, and Clonorchis sinensis are of interest to health professionals, epidemiologists, pharmacologists, and molecular biologists. Recently the transcriptomes of the latter two species were intensively investigated. However our knowledge on molecular biology of O. felineus is scarce. We report the first results of the O. felineus transcriptome analysis. We isolated and annotated a total of 2560 expressed sequence tag (EST) sequences from adult O. felineus (deposited within the database of expressed sequence tags (dbEST), under accession numbers GenBank: JK624271-JK626790, JK006511-JK006547, JK649790-JK649792). Clustering and analysis resulted in the detection of 267 contigs. Of the protein sequences deduced from these, 82% had homologs in the NCBI (nr) protein database and 63% contained conserved domains, allowing the functions to be interpreted using the Gene Ontology terms. Comprehensive analysis of Opisthorchiidae- and Trematoda-specific substitutions within amino acid sequences deduced for the proteins myoglobin, vitelline precursor protein, cathepsin F, and 28kDa glutathione transferase was carried out. The gene set of the 32 ribosomal proteins for the three Opisthorchiidae species with the addition of available Schistosoma and Fasciola orthologs was created and is provided in the supplementary. The orthologous gene set created was used for inferring phylogeny within the Trematoda with special attention to interrelations within the Opisthorchiidae. The phylogenetic analysis revealed a closer relationship between C. sinensis and O. viverrini and some divergence of O. felineus from either O. viverrini or C. sinensis.
Assuntos
Clonorchis sinensis/química , Proteínas de Helminto/química , Opisthorchis/química , Transcriptoma/genética , Sequência de Aminoácidos , Animais , Clonorchis sinensis/classificação , Clonorchis sinensis/genética , Mapeamento de Sequências Contíguas , Cricetinae , Cyprinidae/parasitologia , Cisteína Proteases/química , Cisteína Proteases/genética , Proteínas do Ovo/química , Proteínas do Ovo/genética , Etiquetas de Sequências Expressas/química , Doenças dos Peixes/parasitologia , Glutationa Transferase/química , Glutationa Transferase/genética , Proteínas de Helminto/genética , Mesocricetus , Anotação de Sequência Molecular , Dados de Sequência Molecular , Mioglobina/química , Mioglobina/genética , Opistorquíase/parasitologia , Opistorquíase/veterinária , Opisthorchis/classificação , Opisthorchis/genética , Filogenia , Precursores de Proteínas/química , Precursores de Proteínas/genética , RNA Mensageiro/genética , RNA Mensageiro/isolamento & purificação , Proteínas Ribossômicas/química , Proteínas Ribossômicas/genéticaRESUMO
Experimental preparations based on a DNA vaccine encoding the ESAT-6 antigen of Mycobacterium tuberculosis have been obtained (KpONE6) and studied for immunogenic effects in the murine model. The core of the preparation contains DNA of the recombinant plasmid pONE6 encapsulated within a spermidine-polyglucin conjugate, thereby protecting the DNA vaccine from degradation. KpONE6 induces a proliferative T-cell immune response in mice upon intramuscular immunization.