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Gene sequencing in patients with cancer anamnesis
MARKOVÁ, Iveta
Cancer is the second most common cause of death in the Czech Republic, of which 5-10% are occupied by hereditary cancer syndromes. They are caused by a congenital - hereditary mutation in one of the alleles of the genes, when after the second random intervention in the other allele hereditary cancer develops. It is important to distinguish between hereditary and sporadic carcinomas due to the high risk of inheritance of mutated alleles in the family. The indication may be, for example, the onset of the disease at a young age or the recurrence of the cancer in the family In my work I focused on the analysis and evaluation of data obtained by Sanger sequencing. The aim was to find mutations in the genes mentioned below and to evaluate their pathogenicity by comparison with databases. In the theoretical part of the bachelor thesis I deal with cancer in general and hereditary cancer. I specify the hereditary breast and ovarian cancer syndrome, including genes, that may cause this syndrome - BRCA1, BRCA2, TP53, PTEN, ATM, PALB2, I also deal with Lynch syndrome and the MMR gene system. Last but not least, I describe a familial adenomatous polyposis associated with the APC gene. In the research part I focused on the examination of selected areas of 18 anonymized samples in the gene PALB2 - exon 13 and in the gene BRCA2 - exon 10/4 and exon 11/12. Using the PCR method, I prepared the samples for Sanger sequencing, which then took place in GenSeq s.r.o. In the last part of my work I deal with the analysis and evaluation of the results using the BioEdit program and the NCBI database. I found a mutation in 5 samples - in 4 it was a deletion of one nucleotide with a conflicting interpretation of pathogenicity, the last mutation was pathogenic - causes hereditary breast and ovarian cancer syndrome, it was a nucleotide duplication.
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Autoimmune and lymphoproliferative diseases: associations and common mechanisms
Dobiášová, Alena ; Daňková, Pavlína (advisor) ; Hušáková, Markéta (referee)
Autoimmune and lymphoproliferative diseases share some etiologic mechanisms. The origin of the diseases is complicated process that involves an accumulation of hereditary and somatic mutations in a hematopoetic cell, which thanks to changed activity overcomes different growth and survival control checkpoints. Such mutations are for example those located in genes coding for transcription factors, apoptotic signaling molecules, costimulatory molecules and secreted exctracellular molecules. All these molecules influence the balance between survival and programmed cell death. Their dysregulated expression enables the cell to overcome defensive mechanisms of the immune system. Therefore, autoimmune and malignant cells are able to survive though, under usual circumstances, they would be selected. The main aim of this work is to shed the light on the influence of the dysregulated expression of the particular molecules on the origin of autoimmune and lymphoproliferative diseases. Key words: autoimmune ilnesess, lymphoproliferative diseases, etiology, AIRE, c-MYC, TP53, FOXP3, Fas, PTEN, Bim, CTLA-4, CD5, CD30, CD40/CD40L, BAFF, α-taxilin, IL- 10.
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Cloning and characterisation of selected Class II formins
Stillerová, Lenka ; Cvrčková, Fatima (advisor) ; Rösel, Daniel (referee)
Formins are proteins involved in regulation and construction of actin filaments of eucaryotic organism. They parcipitate in regulating cytokinesis, polar tip growth, and thus participate in development of whole organisms. There are 2 classes of formins in Arabidopsis thaliana. Both classes include FH1 and FH2 domains (formin homology 1 a 2). Class I formins have N-terminal transmembrane domain, unlike class II formins. Some formins of class II have a N-terminal PTEN domain (Phosphatase and Tensin Homolog). Sequence analyses predicted membrane binding via phosphatase or C2 subdomain of PTEN. This thesis was focused on the formin AtFH14, specifically its PTEN domain. Based on predicted sequence, a DNA fragment encoding the PTEN domain was amplified, sequenced and cloned to destination vectors for YFP and EOS phusions. Marked protein was visualized by transient expression in Nicotiana benthamiana. Stably transformed Arabidopsis lines were prepared for stably expression of protein. The tagged protein was localized in cortical cytoplasm, cytoplasmatical strands, probably in nuclear membrane or perinuclear cytoplasm, as well as in peculiar "folicle-like" structures that might be due to binding of PTEN at the periphery of some membrane organelles. Also were seen filament structures, maybe caused by PTEN binding...
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