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Ověření dědičnosti markerů mikrosatelitového panelu u velbloudů
Grygarová, Tereza
The diploma thesis is focused on the verification of inheritance of selected microsatellites from the microsatellite panel of the MHC locus in camels. The theoretical part deals with the characteristics of the genus Camelus. Special focus is given to adaptation mechanisms allowing camels to survive in conditions that are inhospitable for other livestock species. Microsatellites which are also described became an excellent choice not only in parentage analysis. The practical part was based on fragment analysis in genetic analyser ABI PRISM 3500, which preceded DNA isolation, multiplex PCR amplification and gel electrophoresis. Using GeneMapper 5 software, amplified fragment sizes were calculated and mendelian segregation verified. For assembly microsatellite panel four microsatellite markers were selected, M29_I, M35_II, M41_III a M42_III. The least polymorphic was microsatellite M29_I of the MHC I region and the most polymorphic was microsatellite M42_III of the MHC III region. In general, microsatellites from the microsatellite panel can be considered reliable for verifying the origin of the alleles and can be further used to reveal polymorphism of the MHC region of camels.
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Early cancerogenesis in head and neck squamous cell carcinoma.
Kastner, Jan ; Betka, Jan (advisor) ; Salzman, Richard (referee) ; Smilek, Pavel (referee)
Squamous cell carcinoma is the most frequent malignant tumour of head and neck. Prognostic and predictive information as an individual imprint of molecular-genetic analysis of HNSCC will help to determine the best indivicual treatment. And in case of surgical appraoch the optimal resection with adequate quality of life and long-term survival. Study of early cancerogenesis in our project is based on knowledge, that histological normal mucosa next to tumor shows preneoplastic molecular alterations. Molecular genetic changes in a histological normal mucosa harbouring a tumor may play a principal role in revealing of early cancerogenesis process. Molecular-genetic analysis of cancerogenesis in HNSCC reveals prognostic and predictive factors, which are necessary for evaluation and decission for the best individual treatment. This is the concept of tailored medicine The text summarizes current knowledge of early cancerogenesis in HNSCC and presents molecular-biological trends, which are necessary to discover details of early cancerogenesis and thus to get a tool for better detection as well as treatment of malignant disease. The study is based on fragment analysis of microsatelites lesions in tumor tissue in comparison to adjacent mucosa and the healthy mucosa. Results show significant molecular-biological...
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Population biology of the pine needle pathogen Lecanosticta acicola (Thüm.) Syd. (Capnodiales, Ascomycota)
Janoušek, Josef
Lecanosticta acicola is a heterothallic ascomycete that causes brown spot needle blight (BSNB) on native and non-native Pinus spp. in many regions of the world. The aim of this thesis was to elucidate the origin of L. acicola populations in Europe and consider the reproductive mode of the pathogen in affected areas. In order to study the population genetics of L. acicola, eleven polymorphic microsatellite markers were developed. In addition, mating type markers that amplify both mating type idiomorphs (MAT1-1 and MAT1-2) were designed and the protocols for their applications were optimised. Collections of diseased material were obtained from 17 host species in Asia, Europe and America. In total, 201 isolates from diseased pine needles were obtained. All isolates were screened with the microsatellite markers and the mating type idiomorph determined with the mating type markers. For 87 individuals, part of the Translation Elongation Factor 1-alfa gene was sequenced. The isolates from Central America were unique, highly diverse and most likely represent a new cryptic species. The isolates from East Asia formed a discrete group. Two distinct populations were identified in both North America and Europe. Approximate Bayesian Computation analyses strongly suggest independent introductions of two populations from North America into Europe. Microsatellite data and mating type distributions showed the presence of sexual reproduction in North America and in Europe. Results from this thesis have showed that European populations of L. acicola originate from North America. This is the first study of L. acicola populations on a global scale.
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Ověřování SSR markerů vhodných pro rozlišování odrůd vybraných zelenin z čeledi Brassicaceae
Sochorová, Jana
In available literature there is no recommended set of microsatellite markers in the improvement of cabbage cultivars (Brassica oleracea conv. capitata) to distinguish F1 hybrids and self-pollinated plants with a parental genotype. That is why microsatellite markers used to other Brassicas had been searched in this work. Tested microsatellite markers were studied for their ability of distinguishing F1 hybrids in seven cabbage cultivars. Two of 23 microsatellite markers were able to distinguish F1 hybrids in six cabbage cultivars. No microsatellite marker was able to distinguish F1 hybrid and parental genotypes of one cultivar, 'A'.
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