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Lecturer(s)
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Pichová Iva, Ing. CSc.
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Sobotka Roman, prof. Ing. Ph.D.
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Course content
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Content of lectures: 1. DNA structure, history, sequencing, plasmids, cloning 2. Recombinant DNA technology, cosmids, BACs, DNA libraries 3. Gene expression in prokaryotes, promoters, expression systems, Escherichia coli 4. Microbial biotechnology, recombinant enzymes, drugs, bioremediation 5. Eukaryotic expression systems, yeast, insect cells 6. Yeast biotechnology 7. Transgenic animals, side-directed mutagenesis 8. Protein folding, design, mutagenesis, industrial enzymes 9 . Transgenic plants, methodology, Bt and HT plants 10. Second generation of transgenic plants, biopharming, regulation, public issues 11. Protein engineering: directed evolution, library preparation, display systems, applications 12. Protein purification, recombinant proteins, protein solubility, chromatographic techniques, protein tagging Content of tutorials: Purification and analysis of recombinant proteins from Escherichia coli
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Learning activities and teaching methods
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Monologic (reading, lecture, briefing), Demonstration, Laboratory
- Preparation for classes
- 100 hours per semester
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Learning outcomes
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Protein engineering, a specific part of genetic engineering, is the process of developing useful or valuable proteins. It is a young discipline, with much research currently dedicated to the understanding of protein folding and protein recognition for protein design principles. Both rational protein design and directed evolution techniques based on random mutagenesis are employed to generate molecules with novel properties. The course focuses on the molecular and genetic tools used to analyze and modify genetic material and to modify organisms to produce desired molecules and proteins. Topics will include sequencing techniques, cloning vectors and hosts, directed mutagenesis, and the manipulation of expression (and its levels) of particular gene products. Special attention will be directed to study biological systems utilized for the large scale production of recombinant autologous or heterologous proteins, focusing on advantages and disadvantages of each system, to allows students to evaluate and solve problems related to the expression of recombinant proteins. Furthermore, during the course, the major applications of genetic engineering in health care, forensics and agriculture will be presented. Historical overview will help students to understand present day technologies. Lectures on protein engineering will focus on preparation of gene libraries, selection of new proteins by display systems (phage, cell and cell-free) and applications (enzymes, antibodies). Students will also learn how to express and purify proteins using available techniques.
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Prerequisites
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Basic knowledge of biochemistry, genetics and cell biology. A minimal laboratory practice - pippeting, preparation of biological buffers, calculation of solution concentrations.
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Assessment methods and criteria
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Student performance assessment
Credit: completion of all laboratory exercises, submission of a laboratory protocols Examination: Written test (min. 50%)
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Recommended literature
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Glick, R., Pasternak, J.J., Patten, C.L. Molecular Biotechnology. 2010. ISBN 978-1-55581-498-4.
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Sheldon J.P., Cochran J.R. Protein Engineering and Design. 2009. ISBN 9781420076585, CRC Press.
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