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Molecular and biochemical characteristics of genetically modified barley plants
KOCKOVÁ, Lucie
Modern agriculture often employs broad-spectrum herbicides combined with herbicide-resistant crops. Usually, this is achieved by altering the crop genom (genetic modification) by inserting of a specific gene coding for resistance to specific herbicide or group of herbicides. Apllying such herbicide thus results in minor crop damage. The resistence of crops against broad-spectrum herbicides depends on the genes, which were inserted into the genom. The gene bar is often used as a resistence -providing element. It mediates resistance against a broad spectrum of herbicides, such as glufosinate and Bialahops. For the selection of transformants and preliminary assessment of the target transgen expression level a suitable marker gene is simultaneously inserted. For this purpose, gene for bacterial ??glucuronidase (GUS) is often used. It is considered to be one of the most frequently used reporter systems for the assessment of transgen expression and also allows to analyze the expression on the tissue, cell and whole cell organelles levels. In this diploma thesis the PCR method and histochemical detection of the enzyme glucuronidase presence were used to detect and evaluate transgenic plants of cv. Golden Promise spring barley, modified by genes bar and gus. The presence of gus gene was determined in different parts of plants.
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Methods of DNA extraction from the fresh papaya fruit and from the candied
Ovesná, Jaroslava ; Hodek, Jan
Aim the work is to provide a working procedure for extraction of amplifiable DNA form papaya fruits (Carica papaya), which are a poor source of DNA. Methods are optimised for DNA extraction from papaya fruits and stones and from candied papaya- these both were predicted as suitable commodities for an eventually screening of non-approved GM papaya in the market of Czech Republic. The methodology was prepared on basis of demand of reference laboratories of public service, which are involved in GMO analyses and also as a flow-work of Methodics for Detection of GM papaya 55-1 and 63-1 (Hodek, Ovesná, Pavlátová 2008) and scientific publication Detection of transgenic papaya lines: extraction protocol optimisation and verification of DNA quality by PCR assay (Ovesná, Hodek 2009).
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Genotyping of \kur{Giardia intestinalis} isolates
ŠRÁMOVÁ, Eliška
The aim of this work was assemble isolates of Giardia intestinalis from humans and other mammals. Stools samples were examined for presence of cysts by concentration settling method. Consequently sequencing of 532 bp parts of the TPI gene after previous amplification by the nested PCR was performed. In vitro cultures of selected isolates were established using experimental model hosts, gerbils.
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Blastocystis in domestic birds
HRDLIČKOVÁ, Jaroslava
During 2010, faeces samples for parasitology research oriented on Blastocystis were collected from an anonymous farm. A total of 55 samples were collected (of them for 10 faeces samples from hens, ducks and geese and 25 from pigeons). The samples were cultured in Dobell-Leidlaw medium and viewed by light microscopy. However, this method led to only two foundings of Blastocystis (one from hen, the other from pigeon). Thus, PCR and nested-PCR with specific primers were later used for better detection of Blastocystis. The samples for PCR detection were not collected from the aforementioned farm, but they originated from a collection of isolated DNA samples that was available on the Parasitology institute of AS CR. The results of PCR were checked after electrophoresis and verified by sequenation. The obtained sequences of bird-isolated Blastocystis were phylogenetically analysed and as described to subtype 7.
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Isolation of the antimicrobial peptide gene (defensin) from the hard tick \kur{Ixodes ricinus}, challenged by the pathogen infection
SKLADANÁ, Veronika
Antimicrobial peptides are major components of the innate immune response of epithelial cells. In hematophagous organisms, which acts as vectors of parasitic diseases, in particular Lyme borreliosis, the gut pathogen induce the expression of defensin, that provide the first barier of host defence. This raises the posibility that defensin may play a key role in the development of parasitic infection. The gene expressed in midgut was isolated from cDNA of the hard tick, Ixodes ricinus. The gene is coding the protein, that is produced and secreted during tick infection, and is known as defensin. Expression of the gene for defensin (224 bp) was induced by the pathogen. The gene was cloned into bacterial expression system.
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