RESUMEN
Insect mitochondrial genomes (mitogenomes) are of great interest in exploring molecular evolution, phylogenetics, and biogeography. So far, only 12 mitogenomes of the leafhopper tribe Idiocerini have been released in GenBank, although the tribe comprises 488 known species including some agricultural, forestry, and horticultural pests. In order to compare and analyze the mitochondrial genome structure of Idiocerini and even the selective pressure of 13 protein-coding genes (PCGs) of the family Cicadellidae, the complete mitogenomes of five species including Nabicerus dentimus, Sahlbergotettix salicicola, Podulmorinus opacus, Podulmorinus consimilis, and a new species of a new genus were determined by next-generation sequencing. The size of the newly determined mitogenomes ranged from 14,733 bp to 15,044 bp, comprising the standard set of 13 PCGs, 22 transfer RNA genes, two ribosomal RNA genes, and a long non-coding control region (CR). The extent of purifying selection presented different pictures in the tribe and the family. The less pronounced genes (0.5 < dN/dS < 1) were nad5 and nad4l in Idiocerin, whereas in the family Cicadellidae including the sequences of Idiocerin, nad1-nad6 and cox1 genes were less pronounced. The codon encoding leucine was the most common in all species, and the codon encoding serine 1 was the most common in all species except for P. opacus. Interestingly, in P. opacus, another of the most common codons is that encoding serine 2. Among the 17 examined species of the Idiocerini, 14 species contained the tandem repeats, and 11 species of them contained the motif "TTATA". These findings will promote research on the structure and evolution of the mitochondrial genome and highlight the need for more mitogenomes in Cicadellidae.
Asunto(s)
Genoma Mitocondrial , Hemípteros , Animales , Genoma Mitocondrial/genética , Hemípteros/genética , Filogenia , Codón/genética , SerinaRESUMEN
A novel surface plasmon resonance (SPR) biosensor using lectin as bioreceptor was developed for the rapid detection of Escherichia coli (E. coli) O157:H7. The selective interaction of lectins with carbohydrate components from bacterial cells surface was used as the recognition principle for the detection. Five types of lectins from Triticum vulgaris, Canavailia ensiformis, Ulex europaeus, Arachis hypogaea, and Maackia amurensis, were employed to evaluate the selectivity of the approach for binding E. coli O157:H7 effectively. A detection limit of 3×10(3) cfu mL(-1) was obtained for determination of E. coli O157:H7 when used the lectin from T. vulgaris as the binding molecule. Furthermore, the proposed biosensor was used to detect E. coli O157:H7 in real food samples. Results showed that the lectin based SPR biosensor was sensitive, reliable and effective for detection of E. coli O157:H7, which hold a great promise in food safety analysis.
Asunto(s)
Técnicas Biosensibles/métodos , Escherichia coli O157/aislamiento & purificación , Contaminación de Alimentos/análisis , Lectinas/química , Proteínas de Plantas/química , Resonancia por Plasmón de Superficie/métodos , Escherichia coli O157/química , Escherichia coli O157/crecimiento & desarrollo , Microbiología de Alimentos , Unión ProteicaRESUMEN
A surface plasmon resonance (SPR) immunosensor was developed for the detection of E. coli O157:H7 by means of a new subtractive inhibition assay. In the subtractive inhibition assay, E. coli O157:H7 cells and goat polyclonal antibodies for E. coli O157:H7 were incubated for a short of time, and then the E. coli O157:H7 cells which bound antibodies were removed by a stepwise centrifugation process. The remaining free unbound antibodies were detected through interaction with rabbit anti-goat IgG polyclonal antibodies immobilized on the sensor chip using a BIAcore 3000 biosensor. The results showed that the signal was inversely correlated with the concentration of E. coli O157:H7 cells in a range from 3.0 × 10(4) to 3.0 × 10(8) cfu/mL with a detection limit of 3.0 × 10(4) cfu/mL. Compared with direct SPR by immobilizing antibodies on the chip surface to capture the bacterial cells and ELISA for E. coli O157:H7 (detection limit: both 3.0 × 10(5) cfu/mL in this paper), the detection limit of subtractive inhibition assay method was reduced by one order of magnitude. The method simplifies bacterial cell detection to protein-protein interaction, which has the potential for providing a practical alternative for the monitoring of E. coli O157:H7 and other pathogens.
Asunto(s)
Bioensayo/métodos , Escherichia coli O157/aislamiento & purificación , Resonancia por Plasmón de Superficie/métodos , Animales , Anticuerpos Antibacterianos/inmunología , Centrifugación , Ensayo de Inmunoadsorción Enzimática , Escherichia coli O157/citología , Escherichia coli O157/inmunología , Proteínas Inmovilizadas/inmunología , Reproducibilidad de los Resultados , Resonancia por Plasmón de Superficie/instrumentaciónRESUMEN
A biosensor based on surface plasmon resonance (SPR) was developed for rapid detection of Escherichia coli O157 : H7. BIACORE 3000 SPR instrument and a dextran-modified sensor chip (CM5) were used. After activation with EDC/NHS, anti-E. coli O157 : H7 antibody was immobilized on the gold surface of the SPR sensor, and then following ethanolamine was injected, and the chip was ready for E. coli O157 : H7. Regeneration was achieved using NaOH in order to detect several samples. The limit of detection was found to be 3 x 10(5) CFU x mL(-1) for E. coli O157 : H7, and the change of RU was linearly correlated with the concentration of E. coli O157 : H7 cells (R2 = 0.99). The detection time ranged from 5 to 7 min, and the result of regeneration was effective which allowed the chip to be reused for more than 50 samples. This method is convenient, and stable, and shows potentials for applications in food areas.
