RESUMO
Objective: To screen for the optimal qPCR primers for Echinococcus multilocularis apomucin gene (Em-apo) and analyze Em-apo expression. Methods: Primers were designed based on 4 Em-apo sequences from GeneDB. Primer specificity and PCR efficiency were determined, based on which the optimal primer pairs were selected. Alterations of Em-apo expression in 1 000 E. multilocularis protoscoleces treated with albendazoleï¼5 µg/mlï¼ and insulinï¼100 ng/mlï¼ were separately assessed using the selected primers. DMSO used in albendazole dilution and in PBS insulin dilution were used as the control. Results: Specific primers for Em-apo-1, Em-apo-2/3, Em-apo-4 and actin were selected. qPCR melting curves revealed a single peak for each primer pair and an amplification efficiency from 95% to 101%. The qPCR showed increased expression of Em-apo-1ï¼1.51±0.27ï¼, Em-apo-2/3 ï¼1.39±0.30ï¼ and Em-apo-4ï¼1.14±0.18ï¼ after albendazole treatment in comparison to the DMSO controlï¼1.00ï¼ï¼P>0.05 among the three genesï¼; and an unaltered Em-apo-1 expression, slightly decreased Em-apo-4 expression, and significantly decreased Em-apo-2/3 expressionï¼0.73±0.09ï¼ after insulin treatment in comparison to the PBS control (P>0.05 among the three genesï¼. Conclusion: The selected specific primers for Em-apo genes can be used to analyze the gene expression by qPCR. Treatment with albendazole and insulin show certain effects on the expression of Em-apo genes in E. multilocularis protoscoleces.
Assuntos
Echinococcus multilocularis , Albendazol , Animais , Equinococose , Mucinas Gástricas , Reação em Cadeia da Polimerase em Tempo RealRESUMO
The drill (Mandrillus leucophaeus) is a primate of the family Cercopithecidae (Old World monkeys). Drills are among Africa's most endangered mammals, and are listed by the IUCN as the highest conservation priority of all African primates and are used as a model for cytomegalovirus vaccine and antiviral development. Here, we describe the complete mitochondrial genome (mitogenome) sequence of M. leucophaeus. The genome is 16 547 bp in length, comprising 13 protein-coding genes, 22 tRNAs, 2 rRNAs and a major non-coding region. The gene content and order is in accord with the common vertebrate form. All PCGs share the start codon ATG except ND2(ATC), ND3(ATT) and ND5(ATA). ND1, ND2 and ATP8 use ATG as the end codon. COX1, COX2, ATP6, ND4L and ND5 employ ATT as the end codon. The other five PCGs share the end codon T--. Phylogenic tree was constructed based on the complete mitogenome of M. leucophaeus and 12 closely related species to estimate their phylogenic relationship. We present an important genetic resource for the family Cercopithecidae in general.
Assuntos
Genes Mitocondriais , Genoma Mitocondrial , Mandrillus/genética , Filogenia , Animais , Sequência de Bases , Códon , DNA Mitocondrial , Espécies em Perigo de Extinção , Feminino , Ordem dos Genes , Genômica , Análise de Sequência de DNARESUMO
In this study, the complete mitochondrial genome of Dorcadia ioffi was determined. The mitogenome is 16,785 bp in length and contains 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes, and 1 control region. The nucleotide composition of the D. ioffi mitogenome was A: 40.10%, T: 40.61%, G: 7.74%, C: 11.55%. The A + T content is 80.71%, showing strong AT skew. Phylogenetic analysis indicated that Siphonapteran may have sister relationship with Dipteran.