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Efekt polynenasycených mastných kyselin n-3 ve výživě potkana na expresi vybraného genu
Zamazalová, Nikola
The aim of my thesis on topic The effect of n-3 polyunsaturated fatty acids in the diet of rats on expression of selected genes was to investigate the effect of eicosapentaenoic acid (EPA) and docosapentaenoic acid (DHA) on the expression of genes which encode GPR120, AdipoR1 and AdipoR2 receptors in relation to suppress low-grade chronic inflammation in organism to reduce the risk of atherosclerosis by dietary intervention in rats. Rats were fed by a mixture MYPO with 6 % safflower oil (diet S), 6 % fish oil (diet F) or 6 % of oil from algae Schizochytrium (diet A). Gene expression was measured by quantitative real-time PCR method and the results were evaluated by using the software qbase + (Biogazelle NV). Relative expression of GPR120 gene was in F diet 88 % (P > 0,05), in A diet 93 % (P > 0,05) in comparison with control group (100 %). Relative expression of ADIPOR1 gene was in F diets and A diet 82 % (P < 0,05) in comparison with the control group. For ADIPOR2 gene relative expression was 71 % (P < 0,05) in diet F and 68 % (P < 0,05) in diet A. The results were contrary to our hypothesis. However, they exactly match the results of other studies in the available literature. It would be appropriate to carry out further studies on this issue.
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Analysis of intron sequence variability
Kukačková, Hana ; Provazník, Ivo (referee) ; Kubicová, Vladimíra (advisor)
This bachelor thesis reviews genetic expression, the main characteristics of introns, their importance, and the methodology of their recognition. Practical part focuses on intron variability analysis and conservative sequences searching. In the beginning, there is a structure of nucleic acids with their chemical and physical characteristics reviewed, follow by gene expression description with all steps – transcription, posttranscriptional splicing and translation. Afterwards, there is an analysis of introns importance, introns evolution theories and similarities among different organisms mentioned, follow by some introns recognition methods. Practical part is focused on the initial and terminal regions of introns and conservative dinucleotide sequences verifying. The thesis studies intron – exon and exon – intron boundaries. The last part tries to suggest new potentially conservative sequences – patterns, search for them and check their conservativeness.
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DNA Microarrays Data Analysis
Hebelka, Tomáš ; Jaša, Petr (referee) ; Burgetová, Ivana (advisor)
This work concerns with data analysis of DNA microarrays by using cluster analysis. It explains biological terms - gene expression and DNA microarray. Next, it contains mathematical and informatical description of clustering methods and describes a way to apply these methods to microarrays data. Next, the work contains implementation's detail of clustering methods k-means, DBSCAN and introduces an original clustering algorithm Strom++. Then, description of implementation and application manual follow. Finally, accomplished results are evaluated.
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Vliv PUFA n-3 na expresi genů kódujících proteiny řídící homeostázu cholesterolu
Hyblerová, Dagmar
The aim of this study was to confirm that the polyunsaturated fatty acids n-3 (n-3 PUFA) have a positive effect on plasma lipids. These acids can reduce cholesterol by increasing gene expression Insig-1 while decreasing the expression of genes encoding Hmgcr and Ldlr. We tested in experimental rats, which were added to the feed mixture of 6 % safflower oil , 6 % fish oil or 6 % of the oil from the algae Schizochytrium. Relative gene expression was Insig-1 in the test group with addition of fish oil to 120% of controls (P<0.05) and in the group with addition of oils from algae Schizochytrium the relative expression of 170 % of control (P<0.05). These results confirm our hypothesis, only a part, as the relative expression of the gene and Hmgcr and Ldlr was in the test group with addition of fish oil 103% (P>0.05) and 101 % of control (P>0.05) and in the group with addition of oils from algae Schizochytrium the relative expression of 117% (P>0.05) and 156 % (P>0.05) compared to control. Thus, to reduce the relative expression of these genes did not. However, we have shown that n-3 PUFA contribute to a reduction in plasma cholesterol and in this case up to 20 % of control. The concentration of cholesterol in the group with addition of safflower oil was 1.35 mmol.l-1, the group with the addition of fish oil 0.98 mmol.l-1.
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Analýza exprese genů pro růst a vývoj svaloviny u prasat, studium variability genu IL18 a jeho asociace s užitkovými znaky
Chalupová, Pavla
The work builds on the results of microarray analysis and deals with the analysis of differential expression of muscle tissue in pigs. Using quantitative real-time RT-PCR the expression of 12 genes (ANXA1, ANXA2, BTG2, CSRP2, DAB2, EMP3, HBEGF, IL18, PRKAR1A, SGCD, SORT1, TGFB2) involved in the development and growth of muscle in pigs was validated. Differential expression of two muscles (m. longissimus lumborum et thoracis and m. biceps femoris) between Wild Boar and Czech Large White, which differ significantly in growth, was determined. All genes were overexpressed in Czech Large White. In order to resequencing and polymorphism detection, IL18 gene, encoding proinflammatory cytokine was selected. IL18 was previously associated with lipid metabolism, insulin-resistance and skeletal muscle function. In fragments covering exon 2, 4 and 5 a total of 40 polymorphisms were found. PCR-RFLP methodology was designed for two polymorphisms: ENSSSCG00000015037:g.11237G>A in exon 2 (MspI) and g.19480G>A in intron 5--6 (VspI) and the association analysis with the performance and meat quality traits in selected populations in pigs (n = 344 and 669) was evaluated. IL18 was associated with growth, muscle formation, fat deposition and meat quality traits. IL18 could be potentially useful for MAS in pigs.
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Gene expression reporters in \kur{Drosophila melanogaster}.
ŠTROS, Jiří
Drosophila melanogaster is a widely used model organism in genetic research and in a number of other disciplines associated with medical and biotechnological issues. The first part of this thesis presents a review of some basic genetic tools and gene expression reporters used in D. melanogaster research with emphasis on the use of GFP reporter. The second part presents a description and results of my experimental work aimed at the reporter construct consisting of adenosine deaminase gene (ADGF-A) and GFP marker. When using gene expression reporters such as GFP, it is important to know whether the presence of reporter does not affect the studied process. The experiment described in this study tested whether the fusion protein consisting of GFP and adenosine deaminase is fully functional enzyme or whether the enzyme activity is influenced by the presence of fluorescent tag. Results of this work support the usefulness of using the fusion construct as a gene expression reporter.
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The role of genetic background of the host on the pathogenesis of tick-borne encephalitis
PALUS, Martin
We examined the influence of the host genetic background on the pathogenesis of tick-borne encephalitis. We determined virus titers in organs and serum in different time intervals post-infection for different ways of inoculation. We also stated mean survival times and antibody production in different strains of mice infected with tick-borne encephalitis virus. Moreover, differences in expression of immunologicaly important genes in brains of infected mice were compared.
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