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The Interaction of Borrelia Outer Surface Proteins with Tick Salivary Proteins
PAKTAN, Aylin
The aim of this thesis was to investigate the interaction of the salivary gland protein, Salp15, in relation to the outer surface protein C, OspC, using in silico analysis, as well as trying to amplify Salp15 from different tick species found in same geographical areas and observing the growth of Borrelia in the midgut and salivary glands of these different tick species in vitro.
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Porovnání promotorů pro využití v klíštěcím (\kur{Ixodes}) expresním systému
MUSTACOVÁ, Johana
Gene manipulation can be a convenient tool for tick control, however functional procedure for gene manipulation of ticks was not determined yet. The creation of plasmid vectors with high ability of gene expression driven by functional promoters is crucial for genetically modified ticks. To investigate effective tools for tick genes manipulation within the scope of this master thesis, transfection procedures for various types of Ixodes tick cell lines were optimized. As well as expression vectors for use in tick cells were tested. For this purpose, expression plasmid vectors containing luciferase reporter genes driven by eukaryotic and viral promoters were used.
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Production and functional characterization of tick salivary protease inhibitors
KOTÁL, Jan
Two cysteine and two serine protease inhibitors from a tick Ixodes ricinus saliva were overexpressed using a prokaryotic overexpression system and refolded to their native state. Both cysteine protease inhibitors were tested as potential antigens for an anti-tick vaccine showing no effect on tick feeding or reproduction. Various immunological methods were employed to test the potential immunomodulatory function of these proteins without success.
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IrAM9 - a member of a thioester-containing protein family from the hard tick \kur{Ixodes ricinus}
RATHNER, Petr
The thioester-containing proteins (proteins of ?2-macroglobulin family) are constituents of innate immunity, which comprise (i) universal protease inhibitors of ?2M type, (ii) C3, C4, C5 components of the complement system, (iii) insect thioester-contaning proteins (iTEPs) and (iv) macroglobulin complement related proteins (MCR). In this work, the partial structure of the thioester-containing protein from the hard tick Ixodes ricinus (IrAM9) which belong to the MCR was determined by sequencing and cloning of PCR products. The IrAM9 message is expressed in the ovaries and salivary glands of partially engorged females. Received recombinant protein was used to get polyclonal serum which was obtained by repeated rabbit immunization. The obtained antibodies were used in an attempt to detect the native protein in tick tissues by immunoblotting method.
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