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Lecturer(s)
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Fellner Martin, prof. RNDr. Ph.D.
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Nožková Vladimíra, Mgr. Ph.D.
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Husičková Alexandra, Mgr. Ph.D.
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Plíhal Ondřej, RNDr. Ph.D.
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Balarynová Jana, Mgr. Ph.D.
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Karady Michal, Mgr. Ph.D.
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Course content
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1) Basics of plant photomorphogenesis Difference between skotomorphogenesis and photomorphogenesis, plant photoreceptors, effect of blue and red light on plant growth and development, different growth responses of hypocotyls and roots to light, importance of photomorphogenic mutants in study of molecular mechanisms of light sensing and its signaling. 2) Gasometric methods Measurement of gas exchange and photosynthetic parameters. Principle of gasometric parameters (principle of IRGA, different types of measuring systems). Parameters of measured by gasometric systems. Theory of inductive, light and CO2 photosynthetic curves. 3) Basics of chlorophyll fluorescence in vivo Analysis of very fast changes at induction of chlorophyll fluorescence. Kinetics of extinguishing of chlorophyl fluorescence and most often used parameters of both named methods. 4) Basic methods of determination of simple chemicals in biological material, methods of extraction and detection focused on plant material 5) Basics of fluorescent and electron microscopy, basics of cloning and gene engineering, methods of quantification of gene expression Epifluorescent and confocal microscopy, methods of cytosolic staining, immunofluorescent labeling, electron microscopy in biology, GFP technology, molecular cloning, cloning vectors, genomic libraries, heterologous expression systems, gateway technology, methods of transgenosis in plants and new methods of sequencing 6) Quantitative PCR (qPCR) Basic principles of PCR methods, principle of quantitative PCR, strategy o primer design for qPCR, determination of primer efficiency, reagent, primer efficiency, reagents, choice of housekeeping genes, determination of their stability, quantitative gene expression 7) Electrophoretic methods Principles of electrophoresis, types of electrophoresis (agarose, acrylamid, vertical, horizontal), separation of nucleic acids, protein separation (SDS PAGE, native PAGE)
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Learning activities and teaching methods
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Monologic Lecture(Interpretation, Training), Dialogic Lecture (Discussion, Dialog, Brainstorming)
- Attendace
- 24 hours per semester
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Learning outcomes
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To introduce students with selected topics from plant molecular biology supplied with current scientific information from the particular area.
After completing the course, the student should be able to: - Explain basics of plant photomorphogenesis - Describe gasometric methods - Explain basics of chlorophyll fluorescence in vivo - Explain basic methods of determination of simple chemicals in biological material - Describe principles of fluorescent and electron microscopy, basics of cloning and gene engineering, methods of quantification of gene expression - Explain principles of quantitative PCR (qPCR) - Describe electrophoretic methods
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Prerequisites
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Attendance at the course is conditioned by graduation of courses KBB/MBIO1, KBC/BCH, LRR/FZRSB
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Assessment methods and criteria
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Written exam
Oral exam. State final exam.
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Recommended literature
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Arndt C et al. (2012). Native Polyacrylamide Gels. In: Kurien B., Scofield R. (eds) Protein Electrophoresis. Methods in Molecular Biology (Methods and Protocols), vol 869. Humana Press, Totowa, NJ.
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Bustin SA et al. (2009). The MIQE Guidelines: Minimum Information for Publication of Quantitative Real-Time PCR Experiments.
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Dunst S, Tomancak P. (2019). Imaging Flies by Fluorescence Microscopy: Principles, Technologies, and Applications. Genetics 211: 15?34.
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Holčapek M. Hmotnostní spektrometrie v organické analýze. Univerzita Pardubice.
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Jones R et al. (2013). Molecular Life of Plants, Wiley-Blackwell, ASPB.
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Lazár D. (2015). Parameters of photosynthetic partitioning. J. Plant Physiol. 175, 131?147.
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Procházka S, Macháčková I, Krekule J, Šebánek J a kol. (2003). Fyziologie rostlin. Academia. Česká republika.
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Šmarda J. (2008). Metody molekulární biologie. Masarykova univerzita, Brno.
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Von Caemmerer S and Farquhar GD. (1981). Some relationships between the biochemistry of photosynthesis and the gas-exchange of leaves. Planta 153: 376-387.
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