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Studium genetické struktury a diverzity různých populací dravců (Falconiformes)
Bryndová, Marta
The main aim of this thesis was to evaluate the genetic variability in different populations of birds of prey in the Czech Republic. As an alternative source, the feathers were used for the extraction of DNA. The reference species were chosen the Peregrine Falcon (Falco peregrinus) and the Saker Falcon (Falco cherrug), which was also compared with the subpopulation living in Slovakia. Ten microsatellite markers from the literature were tested. Polymorphism of markers varied significantly, locus NVH fp5 was the least polymorphic (PIC = 0.185 F. p; PIC = 0.119 F. ch.). Null alleles were observed in this locus in the Peregrine Falcon population, that is why it should be discarded from the microsatellite panel. The genetic diversity was low among subpopulations of the Peregrine Falcon. FST for the population living in the captivity and living in the wild was 0.025. In the case of the Saker Falcon subpopulation living in Slovakia showed moderate genetic diversity values (0.185 for the subpopulation living in the captivity and 0.126 for the subpopulation living in the wild in the Czech Republic). All subpopulations (except museum specimens of the Saker Falcon) were in Hardy - Weinberg equilibrium. Gene flow was higher among subpopulations of the Peregrine Falcon than the Saker Falcon, where the other population from the different geographical area was also included. 454 sequencing revealed 3 complete mitochondrial DNA sequences of Peregrine Falcons, 2 of Saker Falcons and 2 of Gyrfalcons (Falco rusticolus). The longest sizes were 16,154 bp for the Saker Falcon, 17,239 bp for Gyrfalcon and 17,527 bp for the Peregrine Falcon. Sequence of the Peregrine Falcon was inserted into the Genbank database under accession number JX029991. Whole genome mitochondrial DNA sequences of Saker Falcons and Gyrfalcons have never been published, that is why it will be the part of the new manuscript.
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Studium pozičních kandidátních genů SERPINE1, LDHA a UBL5 ve vztahu k masné užitkovosti prasat
Weisz, Filip
This PhD thesis was focused on study SERPINE1, LDHA and UBL5 as positional candidate genes for meat performance traits in pigs. We detected SNP FN396538:g.566G>A in intron 3 and SNP NM_213910:c.612A>G in exon 3 (Ile159Val) in the SERPINE1 gene, which has been mapped at position 8.4 cM on the linkage map of chromosome 3. Association analyses were conducted on 12th -15th generation of Meishan x Large White (MLW) cross (n = 565) and on European wild boar x Meishan (W x M) F2 population (n = 333). Analyses performed across the entire MLW or in male animals did not show any trait significantly associated with the loci studied. In female animals, both SNPs were significantly associated with loin depth. In the entire W x M population and female animals, SERPINE1 was significantly associated with muscling, growth and fat accretion and in male animals with meat quality traits. In the studied populations, allele effects were in opposite directions, which imply that the SNPs are markers that are in linkage disequilibrium with causative mutations. To search for heterogenity of the LDHA gene we comparatively sequenced cDNA and the 5' flanking region where we detected 8 a 1 SNPs, respectively. We also found 2 different positions of poly(A) sequences in cDNA of LDHA by RACE. Association analyses performed for SNPs GL041338.1:c.49341G>C and FJ865398:c.795A>G did not show any significant association with the studied traits in the MLW population. The significant association of c.49341G>C with growth, fattening traits and muscling in Pietrain x Meishan (P x M) F2 population (n = 316) were found. Association analyses of c.795A>G revealed significant effect of this SNP on growth and fattening traits, fat accretion and muscling in W x M population. The UBL5 gene was studied by inverse PCR and totally 12 polymorphisms were observed. We detected 8 cDNA sequences of various lengths with different transcription start sites of the exon 1 and alternative splicing site between exon 1 and 2 and in 1 sequence also between exon 2 and 3. Alternative splicing between exon 2 and 3 causes shift of the ORF and occurrence of premature stop codon. Association analyses were performed for SNPs FR798948:g.2141C>T and FR798948:g.2778G>A on the MLW and the W x M populations. In the MLW population, SNP g.2141C>T was associated with average daily gain in test and SNP g.2778G>A with back fat thickness. In the W x M population, SNP g.2778G>A was associated with fat accretion, growth and fattening, muscling and SNP g.2141C>T with fat accretion. The SNPs within studied genes that were associated with performance traits are markers in linkage disequilibrium with causative mutation either in genes or in surrounding area.
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Modifikace aplikace pro analýzu lokálních struktur DNA
Zbořil, Jiří
Thesis is focused on a modification and implementation of new features into the existing project, called DNA Analyser. In the first part, there will be analyzing of the actual application structure and tools that are related to it. The second part will be dealing with new implementation in the thesis. This covers the modification of test cases in the application and implementation of new authentication module. There will be discussion related to this solution, as well as a new upgrades and new perspectives to this solution that arise with it.
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Metody izolace a archivace DNA z živočišných tkání
Dratvová, Lenka
n bachelor thesis methods of isolation and archiving of DNA from animal tissues I deal with different samples of animal tissues suitable for efficient isolation of DNA. It selected simple and modern methods of DNA isolation. The work also deals with the possibility of short-term and long-term storage of DNA with a description of the selected archival methods. The method of DNA compared in terms of quality, quantity, and labour, time and monetary perspectives. The individual methods of archiving are also evaluated at the end of the work. The work also includes a practical part, which deals with the isolation of DNA mainly from animal products (milk and milk products, blood, meat and meat products), but also other biological tissues. The procedure and results are presented in chapters "Material and Methods", and "Results and Discussion".
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