Roman Hobza: Genetic Engineering
Semestral course (autumn semester, 2/2)
Mendel University of Agriculture and Forestry, Faculty of Agronomy, branch Plant Biotechnology
- Structure of plant genome
- Complexity of plant genome, nuclear and extranuclear DNA
- Organization of plant chromosome
- Coding and non-coding DNA
- Repetitive DNAsequences, transposons
- General principles of manipulation with nucleic acids
- Isolation of DNA and RNA
- Separation of DNA and RNA: centrifugation, electrophoresis
- Hybridization of nucleic acids
- Enzymes in genetic engineering
- Methods of DNA sequencing
- Polymerase chain reaction (PCR)
- Principle
- Applications: TR-PCR, RAPD, RACE-PCR, AFLP
- Applications: diagnostics, forense analysis
- DNA cloning
- Type of vectors: plasmids, phages, artificial chromosomes
- Techniques of DNA delivery to cells
- Gene transfer into plants
- From DNA discovery to first transgenic plants
- Agrobacterium tumefaciens
- Ti-plasmid and T-DNA
- T-DNA transfer into plant cells
- Transgenic plants
- Resistance against herbicides
- Resistance against herbivorous insects
- Resistance to viruses
- From genes to genomes and back
- Genetic versus physical mapping
- Sequencing of genomes
- Techniques of cytogenetics
- Isolation of chromosomes – flow cytometry and microdissection
- Fluorescence in situ hybridization (FISH)
- Construction of DNA libraries
- cDNA and genomic libraries
- Types of libraries depending on a type of vector
- Application of DNA libraries
- Isolation of genes
- Forward and reverse genetics
- Application of bioinformatics in study of gene structure and function
- Production of proteins in transgenic organisms
- Markers of transgene expression
- Animals as bioreactors
- Modification of physiological processes – golden rice
- Silencing of transgenes
- RNA interference
- siRNA and miRNA
- RNAi and gene knock-out
- Transcriptional and post-transcriptional gene silencing