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Inducible promoters and their use in yeast cell manipulation
Přibáňová, Gabriela ; Palková, Zdena (advisor) ; Vopálenský, Václav (referee)
Promoters which can be regulated by different chemical or physical factors are often used in cell manipulations. This thesis focuses predominantly on promoter systems which use light as an inductor. There are two main approaches to controlling a promoter by light. The first one uses so-called "caged molecules", chemical inducers whose inducing activity is "masked" by a photolabile protecting group. The second approach includes optogenetic systems, which can regulate transcription in cells. These systems are encoded in the DNA of the organism, and light is the only external regulatory stimulus. Photoreceptors that need a specific cofactor (chromophore) are the main components of optogenetic systems. There are several groups of photoreceptors classified by the type of chromophore and photoactivation mechanism. This thesis gives an overview of optogenetic systems used for transcription regulation and focuses on different photoreceptors and induction mechanism used. The systems using photocaged molecules are described as well. Furthermore, the thesis deals with light- systems in yeast as a model organism as well as organism used for biotechnological purposes. Finally, some limitations of light inducible promoters are discussed, including the chromophore type, the wavelength of the light, and the...
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Sublethal effects of the insecticides neonicotinoids on migration abilities of spiders
Přibáňová, Gabriela ; Řezáč, Milan (advisor) ; Korenko, Stanislav (referee)
The purpose for using pesticides is to kill organisms that cause damage (so-called pests) on various crops and thus prevent possible crop losses. Their side effect is a negative impact on non-target organisms. The presented diploma thesis is focused on the effect of neonicotinoids on invertebrates particularly on spiders. Its main goal was to document their influence on behavioral parameters influencing dispersal abilities such as locomotion and tendency to spread by wind (so-called ballooning). The diploma thesis compares the influence of neonicotinoids on model species, Pardosa lugubris (Walckenaer, 1802) and Phylloneta impressa (L. Koch, 1881) with different modes of prey hunting and compares the influence on adult and nymphal stages. Pesticides Actara® 25 WG, Biscaya® 240 OD, Confidor® 200 OD and Mospilan® 20 SP were applied to the spiders under laboratory conditions and their effect was tested at different concentrations and different methods of application. The biggest impact on spiders had Confidor. It clearly had the most significant negative effect on mobility and a tendency to spider wind propagation. The lethal effects were caused by neonicotinoids in nymphal individuals Pardosa lugubris, especially Confidor (even 100% mortality for tarsal application). During an experiment studying the...
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Sublethal effects of the insecticides neonicotinoids on spider's migration and predatory abilities
Přibáňová, Gabriela ; Řezáč, Milan (advisor) ; Korenko, Stanislav (referee)
This research deals with impacts of pesticide substances on spider populations. In this case pesticides of the neonicotinoids classes were tested. Specific neonicotinoids were applied to different of spiders which were collected in various localities and at some stage of development - nymph, adult or subadult individuals. The diploma thesis analyzes and the documents the effects of neonicotinoids to spiders. The results show that some pesticides can have a significant negative effect to locomotion, predation, ballooning, drop or to spiders mortality. Keywords: spider, arthropod, pesticides, insecticides, neonicotinoids, sublethal effects, mortality, EthoVision XT, locomotion, drop, ballooning, predation, Philodromus, Oedothorax, Pardosa, Xerolycosa, Phylloneta
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Inducible promoters and their use in yeast cell manipulation
Přibáňová, Gabriela ; Palková, Zdena (advisor) ; Vopálenský, Václav (referee)
Promoters which can be regulated by different chemical or physical factors are often used in cell manipulations. This thesis focuses predominantly on promoter systems which use light as an inductor. There are two main approaches to controlling a promoter by light. The first one uses so-called "caged molecules", chemical inducers whose inducing activity is "masked" by a photolabile protecting group. The second approach includes optogenetic systems, which can regulate transcription in cells. These systems are encoded in the DNA of the organism, and light is the only external regulatory stimulus. Photoreceptors that need a specific cofactor (chromophore) are the main components of optogenetic systems. There are several groups of photoreceptors classified by the type of chromophore and photoactivation mechanism. This thesis gives an overview of optogenetic systems used for transcription regulation and focuses on different photoreceptors and induction mechanism used. The systems using photocaged molecules are described as well. Furthermore, the thesis deals with light- systems in yeast as a model organism as well as organism used for biotechnological purposes. Finally, some limitations of light inducible promoters are discussed, including the chromophore type, the wavelength of the light, and the...
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