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Nematobacterial complex Heterorhabditis-Photorhabdus in immunity studies

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VOJTEK Libor DOBEŠ Pavel HYRŠL Pavel

Rok publikování 2010
Druh Konferenční abstrakty
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Citace
Popis Entomopathogenic nematodes are obligate insect parasites with the increasing importance as biological control agent. Furthermore, nematodes can be used as unique natural model for infection of numerous insect species combining the simultaneous action of nematodes and their symbiotic bacteria. The bacterial symbionts are essential for successful invasion to the host causing septicaemia inside and digesting host tissues. We use common entomopathogenic nematode Heterorhabditis bacteriophora associated with symbiotic bacteria Photorhabdus luminescens which are capable to kill the insect host within 24-48 hours after infection. Especially in combination with genetically tractable Drosophila melanogaster nematobacterial complex offers ideal tool for studying the insect physiology and immunity. Symbiotic bacteria can be isolated and separately used for determination of their pathogenity to insect without nematode's influence. Photorhabdus is the only natural bioluminescence genus of soil G- bacteria, therefore it is widely applied for bioluminiscence tests. We use two non-pathogenic subspecies (laumondii and kayaii) of P. luminescens for antibacterial assays based on their bioluminescence ability (e.g. complement, myeloperoxidase and antibacterial activity determination). Except P. luminescens we use genetically modified Escherichia coli K12 that carries Photorhabdus genes for bacterial luciferase (Lux) and its substrate. Bioluminescence reaction is mediated by bacterial enzyme luciferase which catalyses the oxidation of long-chain aldehyde (substrate) and reduces flavin mononucleotide with emission of light. This emission can be directly measured by the luminometer, thus we can assumed bacterial viability. Our research is supported by grant from Grant Agency of Czech Republic (GA206/09/P470).
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