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Lecturer(s)
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Čurn Vladislav, prof. Ing. Ph.D.
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Beran Pavel, Ing. Ph.D.
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Hoštičková Irena, Ing. Ph.D.
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Course content
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Introduction to methods of genetic engineering and recombinant DNA technology. Restriction endonucleases, modification and other enzymes in molecular biology. Plasmids, bacteriophages and bacteria in genetic engineering and molecular biology. Transformation of bacteria. Bakteriophages as vectors in gene library construction. Plasmids - mechanism of replication, incompatibility, copy number, partition. DNA cloning, methods and principles, importance, methods of screening. Antibiotics and resistance. Reporter genes. Gene library, reverse transcription, library screening, protein sequencing, translation in vitro, Western blotting. Methods of in vitro manipulation with DNA and RNA. Nucleic acids isolation and purification, modification and labelling, molecular probes, hybridisation, Southern blotting. DNA sequencing. Sanger and Maxam-Gilbert methods and 454 approaches. RFLP technology. DNA amplification - PCR technology. Molecular markers (RFLP, RAPD, AFLP). Structure of eukaryotic genome, methods of analysis. Basic method of plant genetic engineering. Legal, social and ecological aspects of genetic engineering.
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Learning activities and teaching methods
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Monologic (reading, lecture, briefing)
- Preparation for credit
- 45 hours per semester
- Preparation for exam
- 50 hours per semester
- Semestral paper
- 60 hours per semester
- Class attendance
- 56 hours per semester
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Learning outcomes
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The aim of the course Biotechology of Plant Cell is to clarify the issues of plant molecular biology, manipulation with DNA/RNA and basic methods of genetic engineering. The lectures are primarily focused on mastering the basic methodology used in DNA analysis, recombinant DNA technology and plant transformation. Characterization of Agrobacterium, structure of Ti and Ri plasmid, T-DNA. Transformation of plants using Agrobacterium, direct methods of transformation, somatic hybridization, regeneration and selection of transgenic plants. Signal genes and their determination, evidence of stable T-DNA integration, identification of plants using molecular biology techniques. Stability of expression of introduced genes, practical use of plant transgenesis, current research trends in plant transgenesis.
Students are able to orientate in the field of methods of molecular biology and their application in plants
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Prerequisites
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Standard knowledge of biology and genetics
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Assessment methods and criteria
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Oral examination
Students are able to orientate in the field of methods of molecular biology and their application in plants
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Recommended literature
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Alberts,B. a kol. Základy buněčné biologie. Espero Publishing,2. vydání, 2005.
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Mazura,I. a kol. Speciální metody molekulární biologie. Karolinum, UK Praha, 2001.
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Ondřej,M.,Drobník,J. Transgenoze rostlin. Academia Praha, 2002.
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Reece,R.J. Analysis of genes and genomes. Wiley Publishing, 2004.
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Sambrook,J. a kol. Molecular cloning. New York, CSHL Press, 2001.
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