1. COURSE OBJECTIVES. Understanding chromosomes on molecular level, understanding of chromosome division, interphase chromatin organisation. Techniques of molecular cytogenetics for chromosome/chromatin analyses.
2. LEARNING OUTCOMES. Linking molecular sequencing data with chromosomes. Expression analysis of the tissue level and cell level. Integration of already acquired knowledge of cell and molecular biology and genetics with chromosomes, chromatin and interphase architecture. Techniques of molecular cytogenetics (integration of methods of molecular biology and cytogenetic methods) with special focus on fluorescence in situ hybridisation.
3. CONTENT OF THE COURSE MOLECULAR CYTOGENETICS. 1. INTRODUCTION IN MOLECULAR CYTOGENETICS. Brief historical overview. Techinques of molecular cytogenetics: FISH, GISH and Direct in situ hybridisation (DNA fiber FISH), in situ PCR, PRINS, microdissection, chromosome sorting. 2. GENOME SIZE. Heterogeneity of eukaryotic genomes due to repetitive DNA; Satellite DNA (non classical roles of satellite DNA, satelite DNA and evolution; evolution of satellite DNAs); Minisatelites and microsatellites (use in fingerprinting and genotyping; role in regulation of gene expression); Mobile DNA (types, evolution and roles in eukaryotic genomes); DIstribution of repetitive DNA in chromosomes; Genome size and repetitive DNA (role in animal and plant adaptation). 3. CHROMATIN ORGANISATION. Nucleosome fiber; Solenoid; Higher order chromatin structures. 4. STRUCTURAL REGIONS IN EUKARYOTIC CHROMOSOMES. Sequence and protein composition od centromeric region. Role of centromere region for eukaryotic chromosomes. EPigenetics of centromeric region. Telomeres. Sequence and protein composition of telomeres; role of telomeres in eukaryotic chromosomes; replication of telomeric region; epigenetics of telomeric region. 5. INTERPHASE ARCHITECTURE. Distribution and organisation of chromatin in interphase nucleus; chromatin domains and interchromosome region; distribution and position of chromosomes in different cell types; redistribution of chromatin during cell differentiation. 6. MOLECULAR ASPECTS OF MEIOSIS. Molecular mechanisms during meiosis: finding of homologous chromosomes; homologous recombination (dependent of double strand break and independent of double strand break); synapsis; monopolar and bipolar attachment of chromosome kinetochores; nuclear spindle; cohesines.
PRACTICAL COURSE. Preparation of cytogenetic spreads. Method of fluorescent in situ hybridisation (FISH). Microscopy following FISH and image analysis using computer programmes for image analysis.
SEMINARS
Students choose high quality recent publications (within 5 years) and present in form of Power Point presentation. The professor evaluate the comprehension of the topic, the clarity of presentation and ability of student to connect different topics of the epigenetic material.
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- Molecular Biology of the Gene, Watson JD,Baker TA, Bell SP, Gann A, Levine M, Losick R, Pearson Education Inc., Benjamin Cummings, 2004
Practical in situ Hybridisation, Schwarcher T, Heslop Harrison P, Bios, Scientific Publisher Ltd. 2000
Plant Cytogenetics, Singh RJ, CRC Press London, 2003
Species Evolution: The Role of Chromsome Change, Max King, Cambridge University Press, 1995
Non radioactive in situ hybridisation application manual, Boehringer Mannheim, 1996
PCR Protocols: a Guide to Methods and Applications, Innis MA, Gelfand DH, Sninsky GG, White TJ, Accademic Press, Boston, 1990
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