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MBM


Course: Mol.Biol. of Mutagens, Carcinogens, Cyt.

Department/Abbreviation: KBF/MBM

Year: 2019

Guarantee: 'prof. RNDr. Jana Kašpárková, Ph.D.'

Annotation: Mutagens ansd cancerogens, their definition and biological properties. Mechanism of their interaction with nucleic acids and repair mechanisms of DNA damages. Special attention is given to the experimental methods of studies of DNA interactions.

Course review:
The course will discuss the role of mutagens in living organisms, mechanisms underlying their reactions with nucleic acids and mechanisms of repair of the damage induced by their action. A particular chapter will be devoted to the methods used in the studies of interactions of mutagens with nucleic acids. The issues will be divided in the following way: 1. Chemical compounds as mutagens and caricinogens. Definition of mutations, origin and types of mutations. 2. Basic properties of nucleic acids: Chemical composition, structure and properties of nucleic acids. 3. Mechanism of transfer of genetic information and its alterationsus (replication and transcription), defects in these processes and origin of mutations. 4. Genetic material handling, isolation of nucleic acids and manipulation with nucleic acids. 5. Determination of nucleotide sequence of nucleic acids (endonucleases, exonucleases, restriction enzymes, chemical /Maxam-Gilbert/ method, the chain-termination /Sanger/ method) how to find out that mutation occurred. 6. Reactions of mutagens, carcinogens and cytostatics: direct reactions, metabolic activation; types of DNA damage. 7. Repair of damaged DNA in mammalian cells. I. total repair, excision repair: base excision repair, nucleotide excision repair, mismatch repair. 8. Repair of damaged DNA in mammalian cells. II: repair of double-strand breaks, tolerant repair. 9. DNA damage and cancer. Carcinogenesis. 10. Current approaches in the design of new cytostatics and their development. 11. Cytostatics - types of the most frequently used cytostatics, determination of the target. 12. In vitro cytotoxicity tests. 13. Methods and procedures for determination of mutagenicity of chemical substances and physical agents - tests on molecular, gene and chromosome level. 14. Methods for studies of reactions of mutagens, carcinogens and cytostatics with DNA. I. a) DNA binding, detection of binding and damage, b) determination of sequence specificity of the binding. 15. Methods for studies of reactions of mutagens, carcinogens and cytostatics with DNA. II. a) formation of interstrand cross-links and their quantification, b) alterations if conformation and stability of DNA. 16. Methods for studies of reactions of mutagens, carcinogens and cytostatics with DNA. III. Reactions with cellular proteins and studies of repair processes in vitro and in cells. 17. Electrochemical detection of damage of genetic material.