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Pregnancy proteins - molecular biological and biochemical analysis
Muravská, Alexandra ; Kalousová, Marta (advisor) ; Černá, Marie (referee) ; Průša, Richard (referee)
The aim of this thesis was to establish methods for selected PAPP-A (Pregnancy- Associated Plasma Protein A) gene polymorphisms analysis and to study genetic background of PAPP-A and biochemical background of PAPP-A and PlGF (Placental Growth Factor) in relation to risk pregnancy. Secondly, the aim was to establish method for two-dimensional (2D) electrophoresis of amniotic fluid. Methods for analysis of ten PAPP-A gene polymorphisms were established. These polymorphisms, PAPP-A and PlGF levels were studied in together 165 women in third trimester pregnancies complicated with threatening preterm labor (n=98), preeclampsia (n=35), IUGR (Intrauterine Growth Restriction) (n=34) and ICP (Intrahepatic Cholestasis of Pregnancy) (n=15). 114 healthy pregnant women served as controls. The method for 2D electrophoresis of amniotic fluid was established. Preeclamptic patients had significantly higher frequency of TT genotype of Cys327Cys (C/T) PAPP-A gene polymorphism compared to controls. Patients with ICP had increased serum levels of PAPP-A compared to controls, in patients with threatening preterm labor PAPP-A levels were rather decreased. PlGF levels did not differ from control group in patients with ICP and threatening preterm labor. Positive correlation was found between PAPP-A and PlGF in group of...
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Signal processing based methods for genome assembly refinement
Jugas, Robin ; Provazník, Ivo (referee) ; Sedlář, Karel (advisor)
The diploma thesis deals with sequencing methods and genome assembly methods including usage of numerical representations. The theoretical part of thesis describes the history of DNA research, generations of sequencing methods, the assembly methods themselves and definiton of numerical representations. Numerical represenatations serve to convert character form of DNA to numerical form and so allow to use digital signal processing methods. There is an algorithm for genome assembly using numerical represenatation proposed in thesis, which is later tested at sequence data.
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Genotyping of bacterial genus \kur{Xanthomonas} pathogenic for tomato and pepper by multilocus sequencing.
STEHLÍKOVÁ, Dagmar
The subject of this bachelor thesis is to estimate the degree of relationship of 20 collected isolated samples of Xanthomonas genus causing bacterial spot of tomatoes and peppers. To assess the relationship, the method of multilocus sequence analysis has been used (MLSA). Sequences of genes atpD, rpoD, gyrB, glnA and efP were found and analyzed using MEGA software version 5.1. The result was evaluated using a dendrogram according to which the isolates were divided into two clusters. The first cluster includes isolated of Xanthomonas euvesicatoria belonging to group A. The second cluster includes isolates of the species Xanthomonas vesicatoria belonging to group B. The results obtained in this work are consistent with the present classification of the xanhtomonads causing bacterial spot of tomatoes and peppers.
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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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Identification and sequencing genom of a new virus infecting lucerne
BEČKOVÁ, Martina
Samples of lucerne plants characteristic with local necrosic lesions, leave malformation and yellow spots on leaves were investigated with transmission electron microscopy. Virus particles observed there were filamentous ones of 600 to 700 nm long. Nucleic acid was isolated, transcribed and amplified using PCR. Genus-specific primers were designed based on reverse genetics from the highly conserved genes for carlaviruses, potexviruses and potyviruses. Successful amplification with carlavirus-specific primers, sequencing and comparison with sequences in GenBank database revealed presence of a carlavirus. This was later identified by nucleotide sequence comparison as a new isolate V4 of Alfalfa latent virus. Specific primers for isolate V4 were designed in a coat protein position. Half of the genom of this virus was obtained with PCR and PCR modified amplifications and compared with sequences of Alfalfa latent virus and Pea streak virus from GenBank.
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