RESUMEN
OBJECTIVE: To determine the phylogenetic position of Schistosoma sinensium in the genus Schistosoma using mitochondrial cytochrome C oxidase 1 (CO1) and NADH dehydrogenase 1(ND1) as molecular markers. METHODS: The genomic DNA of adult worms were extracted by the GNT-K method. The target regions were amplified by PCR using specific primers. The PCR products were purified before ligation into the plasmid Zero-Blunt. Recombinant plasmids were amplified in E. coli, extracted and purified using routine methods and then sequenced using M13 primers (F/R) on a Licor long-read auto-sequencer. Sequences of related schistosomes were retrieved from GenBank and aligned with our data in the sequence editor ESEE. Gene trees were constructed in PHYLIP and MEGA using both maximum parsimony and neighbor-joining methods. For parsimony analysis, all characters were treated as unordered and with equal weights. At least 3,000 cycles of bootstrapping were carried out. For analysis in MEGA, all gap columns were deleted. The third position of codon was included. RESULTS: The nucleotide and amino acid sequences of CO1 and ND1 of S. sinensium were obtained. CONCLUSION: The phylogenetic trees from these molecular data suggested that S. sinensium belongs to the Asian schistosome group, and the results coincided with the previous rDNA (ITS2 & LSU) analysis results.
Asunto(s)
Genes Mitocondriales , Schistosoma/clasificación , Animales , Complejo IV de Transporte de Electrones/química , Complejo IV de Transporte de Electrones/genética , NADH Deshidrogenasa/química , NADH Deshidrogenasa/genética , Filogenia , Schistosoma/genéticaRESUMEN
OBJECTIVE: To study genomic diversity among the isolates of Schistosoma japonicum in China. METHODS: Schistosome adults were collected from different endemic areas. Mitochondrial NADH dehydrogenase 1 (ND1) and cytochrome C oxidase 1 (CO1) gene fragments of the worms were amplified by PCR, cloned into plasmid, and finally sequenced. Program MEGA was used for sequence analysis. RESULTS: The ND1 and CO1 genes amplified from different geographic strains were 476 bp and 1,033 bp, respectively. There are two distinct haplotypes, type I and II, for both ND1 and CO1 nucleotide sequences. The differences between the two types were 4.0% and 3.4%. They are considered to be two different genetypes by the phylogenetic analysis. In individual mitochondrial gene, type I of ND1 was fixedly accompanied with type I of CO1 gene, and type II of ND1 with type II of CO1. CONCLUSION: There are two different genetypes of ND1 and CO1 genes of S. japonicum in China.
Asunto(s)
Complejo IV de Transporte de Electrones/genética , NADH Deshidrogenasa/genética , Schistosoma japonicum/genética , Animales , Secuencia de Bases , China , Genotipo , Haplotipos , Ratones , Mitocondrias/enzimología , Datos de Secuencia Molecular , Filogenia , Schistosoma japonicum/enzimologíaRESUMEN
OBJECTIVE: To classify the taxonomic status of O. cheni in relation to O. turkestanicum var. tuberculata from the mainland of China by comparing their nucleotide sequences of nuclear ribosomal partial large subunit gene (LSU). METHODS: The genomic DNA of adult worms were extracted by the GNT-K method. The target gene was amplified by PCR using specific primers. The PCR products were purified before ligation into the plasmid PCR-blunt (Invitrogen). Recombinant plasmids were amplified in E. coli, extracted and purified using routine methods and then sequenced using M13 primers (F/R) on a Licor long-read auto-sequencer. Sequences of O. turkestanicum was retrieved from GenBank and aligned with our data in BioEdit. RESULTS: The nucleotide sequences of LSU between O. turkestanicum var. tuberculata and O. cheni was 100% identical, and 99.99% identical between O. turkestanicum var. tuberculata and O. turkestanicum. CONCLUSION: This study demonstrated high similarity in LSU nucleotide sequences, and the results do not support O. cheni as an independent species. O. cheni may be a synonym of O. turkestanicum var. tuberculata, and O. turkestanicum var. tuberculata is probably also a synonym of O. turkestanicum.