RESUMO
Toxoplasma gondii, an obligate intracellular protozoan parasite of the phylum Apicomplexa, can infect all warm-blooded vertebrates, including humans, livestock, and marine mammals. The aim of this study was to investigate whether superoxide dismutase (SOD) of T. gondii can be used as a new marker for genetic study or a potential vaccine candidate. The partial genome region of the SOD gene was amplified and sequenced from 10 different T. gondii isolates from different parts of the world, and all the sequences were examined by PCR-RFLP, sequence analysis, and phylogenetic reconstruction. The results showed that partial SOD gene sequences ranged from 1,702 bp to 1,712 bp and A + T contents varied from 50.1% to 51.1% among all examined isolates. Sequence alignment analysis identified total 43 variable nucleotide positions, and these results showed that 97.5% sequence similarity of SOD gene among all examined isolates. Phylogenetic analysis revealed that these SOD sequences were not an effective molecular marker for differential identification of T. gondii strains. The research demonstrated existence of low sequence variation in the SOD gene among T. gondii strains of different genotypes from different hosts and geographical regions.
Assuntos
Animais , Gatos , Humanos , Sequência de Aminoácidos , Sequência de Bases , Variação Genética , Cabras , Dados de Sequência Molecular , Filogenia , Proteínas de Protozoários/química , Alinhamento de Sequência , Ovinos , Superóxido Dismutase/química , Toxoplasma/classificação , Toxoplasmose/parasitologia , Toxoplasmose Animal/parasitologiaRESUMO
Objective:The best operation condition of extraction was found through analyzing the influences of both process of microwave pretreatment and hot wave washing. Methods: A new extraction method of chlorogenic acid was that the dry FLos Lonicerae was humidified with 75% ethanol-water solution at first, then the wet Flos Lonicerae was rapidly vaporized by microwave heating, at last chlorogenic acid was extracted after hot water washed twice, 10 minutes each time. Results: Compared with traditional extraction, the extraction time was about less than 6 times and then extraction ratio was increased by 1%. Conclusion: The method has high rate of extraction and short time of extraction.