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Molecular genetic characterization of vancomycin-resistant enterococci
Bubeníček, Karel ; Rada, Vojtěch (advisor) ; Igor, Igor (referee)
Summary
Objectives and hypothesis: This thesis concerns the study of plasmids of vancomycin-
resistant enterococci isolated from feces of American crows in the years 2012 - 2013 period.
The hypothesis is that, in various environments, there is one or more types of
epidemiologically significant vanA gene-carrying plasmids that are capable of horizontally
spread.
Methods: Based on PFGE method the number and size of plasmids were detected in selected
isolates of vancomycin-resistant E. faecium. Using PCR method the isolates were subjected
to detection of genes of replicases, relaxases and toxin-antitoxin system of plasmid-bound
resistance genes. Using 19 primers were characterized types of Tn1546.
Results: Of the 12 tested vancomycin-resistant isolates of E. faecium the following number
and size of plasmids was proven using PFGE method: 2 isolates contained two plasmids
(17%), 3 isolates contained three plasmids (25 %), 5 isolates contained four plasmids (42 %)
and 2 isolates contained five plasmids (17 %).
All isolates (n = 12) were then subjected to the detection of genes of replicases, relaxases and
toxin-antitoxin system for typing of plasmids from each plasmid families.
RepA_N family of plasmids:
genes characterizing plasmids related to pRUM: rep17 in 11 isolates (92 %),
gene Axe-Txe was detected in 5 isolates (42 %)
genes characterizing plasmids related to pLG1: rep20 in 7 isolates (58 %)
genes characterizing plasmids related to pAD1: relpAD1 gene was detected in one isolate (8 %)
Inc18 family of plasmids:
genes characterizing plasmids related to pIL501: rep1 gene detected in one case (8 %)
genes characterizing plasmids related to pRES25: rep2 gene in 2 isolates (17 %)
genes characterizing plasmids related to pEF1: relpEF1 detected in 11 isolates (92 %)
pHTB family of plasmids:
genes characterizing plasmids related to pHTB: rep22 gene was detected in 4 isolates (33%) and in 2 isolates gene relpHTB was detected (17%)
RCR family of plasmids:
genes characterizing plasmids related to pRI: positive detection of Rep14 gene in 8 isolates (67%) and in 4 isolates relpRI gene was detected
Small theta-replicating plasmids:
genes characterizing plasmids related to pEF418 plasmids: rep18a gene in 2 isolates (17%)
genes characterizing plasmids related to pB82: rep18b gene was detected in one isolate (8%)
genes characterizing plasmids related to pCIZ2: relpCIZ2 gene was detected in 9 isolates tested (75%)
Types of transposon Tn1546
Using the PCR method types of Tn1546 were characterized. In 4 isolates (n = 12; 33 %)
Tn1546 was characterized as a F3 type. In one isolate (8 %) Tn1546 was characterized as a
type F5, in one isolate (8 %) as a type PP-16. In 6 isolates Tn1546 was untypeable. Most
likely these are new, yet unknown types.
Conclusion: This is the first study of plasmids of vancomycin-resistant isolates E. faecium
isolated from feces of American crows. These results emphasize not only a high proportion of
plasmids in individual isolates, but also a high proportion of genes with horizontal transfer.
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Methods for gene prediction in prokaryotic genomes
Nykrýnová, Markéta ; Sedlář, Karel (referee) ; Maděránková, Denisa (advisor)
This bachelor thesis deals with methods of gene prediction in prokaryotic genomes. First part of the thesis introduces prokaryotic cell, its genome, expression of genetic information and methods for gene prediction. Following part describes three software products for gene prediction. Chosen software was tested against specific genome and next chapter presents obtained results. The last part describes program called Gene_finder and its results.
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The importance of predisposition examination of late Alzheimer´s disease using methods of molecular biology
KOPAČKOVÁ, Michaela
Alzheimer's disease is the most common type of dementia. It affects up to 70% of cases from approximately 38 million people worldwide suffering from some type of dementia. Alzheimer's disease usually affects patients over 65 years of age, however, there are exceptions. It is presumed that the number of people with Alzheimer's disease will grow constantly in the future too. Its etiopathogenesis is not yet fully understood, that is why it is important to investigate dementia further, whether in terms of specific environmental and genetic risk factors, or in view of the economic burden for the society. The presented bachelor's thesis summarises findings about Alzheimer's disease, its neurobiology, etiology, diagnostics and genetic predispositions, in particular the ApoE, APP, PSEN1 and PSEN2 genes. The presence of epsilon 4 gene allele for apolipoprotein E provably contributes to the increased risk of the late form of Alzheimer's disease. On the contrary, mutation in APP, PSEN1 and PSEN2 genes are connected with Alzheimer's disease with the early onset of the disease, before 65 years of age. The objective in the experimental part was to manage the practical isolation of DNA from peripheral blood and buccal swab, preparation and execution of PCR reaction, detection of PCR products by gel electrophoresis and independent examination of ApoE gene by a certified CVD StripAssay Kit from ViennaLab company (PentaGen). The work also includes the optimisation of another molecular genetic method for the detection of variant alleles of ApoE gene. This method was carried out by a certified DiaPlexQTM ApoE Kit from SolGent Co., Ltd. The principle of detection in this case is based on real-time PCR.
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Genetická analýza vybrané dědičné choroby u psů
VOLNÁ, Jitka
In my thesis I concentrated on an analysis of biological samples of 60 dogs, which manifested epilepsy. The aim was to find out the relation between genotype of the selected locus and the occurrence of epilepsy. Based on the results of the analysis, I assessed, whether there is a demonstrable relation between the occurrence of epilepsy and genotype in a particular locus. According to the results, no influence of genotype on the age of first epileptic seizure was proved.
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DNA čipy a jejich využití pro molekulární diagnostiku a predikci u onkologických onemocnění
VORÁČKOVÁ, Lucie
Many visible factors produced during carcinogenesis refer to oncological disease, e.g. modified gene's expression or increased number of miRNA. These factors can be diagnosed by analyzing of gene's expression. For this diagnostics, there are used many methods, of whom DNA microarray analysis is the most significant progress in genetic research. A large number of DNA sequences can be detected DNA with microarrays on a single microarray in a relatively short time. This analysis means a positive turnover for the diagnosis and prediction of disease.
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Comparison of genomes by synteny block analysis
Pavel, Tomáš ; Škutková, Helena (referee) ; Maděránková, Denisa (advisor)
The theoretical part of this bachelor thesis is aimed at basics of genetics. Term of gene and mutation are introduced in this section. There are gene and chromosome mutations mentioned and described. Following section is devoted to comparative genomics and especially to synteny. There is described what the synteny actually is and how the synteny arises. The end of the theoretical part of this thesis is about the evolution and there are described ways of sorting permutation vectors. The practical part of this bachelor thesis includes description of developed software. Output of this software is a dot-plot which shows detected synteny blocks. Indexes of these blocks are listed in GUI. The second important output is number of permutation steps. This number determines evolutionary distance between two analysed DNA sequences. The very last section is aimed at analyse of synthetic and real DNA sequences.
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Identifying CpG islands in genomes of eukaryotes
Urbánková, Kateřina ; Provazník, Ivo (referee) ; Maděránková, Denisa (advisor)
Bachelor thesis deals with searching CpG islands in DNA. The theoretical part describes the structure and composition of deoxyribonucleic acid, chromosomes and genes. There is basic information about human and chimpanzees genome and specification of CpG islands. Practical part consists of a program that searches for CpG islands in sequences, it displayes and saves it. This work includes analysis of genes which are common for Homo sapiens, Pan troglodytes, Mus musculus and Bos taurus.
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Bioinformatics of human genome
Kupková, Karolína ; Provazník, Ivo (referee) ; Maděránková, Denisa (advisor)
Bachelor thesis deals with DNA segments containing only adenine and guanine. The theoretical part describes the structure and composition of deoxyribonucleic acid, chromosomes and genes. There are basic informations about chimpanzees and the human genome and conformations of the chains containing adenine with guanine. The practical part consists of a program that searches for the required sections in sequence, it displays and saves it. The thesis includes analysis of genes which are common for humans and chimpanzees, which were analyzed to determine randomness, functionality a preserving of these sections.
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Gene Detection in DNA Sequences
Roubalík, Zbyněk ; Burgetová, Ivana (referee) ; Martínek, Tomáš (advisor)
Gene detection in DNA sequences is one of the most difficult problems, which have been currently solved in bioinformatics. This thesis deals with gene detection in DNA sequences with methods using Hidden Markov Models. It contains a brief description of the fundamental principles of molecular biology, explains how genetic information is stored in DNA sequences, as well as the theoretical basis of the Hidden Markov Models. Further is described subsequent approach in the design of specific Hidden Markov Models for solving the problem of gene detection in DNA sequences. Is designed and implemented application, which uses previously designed Hidden Markov model for gene detection. This application is tested on the real data, results of these tests are discussed in the end of the thesis, as well as the possible extension and continuation of the project.
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