Genetic engineering E BT1502

To familiarize students with knowledge concerning the activities on the genetic material and the ability to work in a research lab in conducting experimental work on nucleic acids (DNA, RNA) and the biological material (cells, tissues, microorganisms, plants, animals); Operating equipment research and control; independently develop professional skills; be prepared to work in laboratories of control and diagnostics; independently conduct the processes of genetic engineering
The lecture is to acquaint students with knowledge of:
The enzymes used in genetic engineering, with focus on restrictases.
The basic genetic techniques in vitro. Isolation of genomic DNA and plasmid DNA and DNA purification. Electrophoresis vertical and horizontal. PCR. Site-directed mutagenesis. Random mutagenesis. Transformation. Sequencing. Hybridization.
Cloning vectors. Plasmid vectors and others.
Genetic recombination. Principle of the method of obtaining genes.
Recombinant DNA expression systems.
Reverse genetics. The search function of genes. A breakdown of the gene - gene disruption. Transcriptional and post-transcriptional silencing RNA (RNAi).
Proteomics. The study of protein function. The study protein-protein interactions.
Laboratory include:
The basic techniques of genetic engineering in vitro.
Isolation of genomic DNA and the plasmid. Isolation of DNA from bacterial cells. Isolation of DNA from human tumor cells and physiological cells.
Isolation of genomic RNA. Isolation of RNA from human tumor cells and physiological cells.
Cutting extracted DNA using the selected restriction enzymes.
The study isolates the purity of DNA and RNA. Long-term storage of biological samples.
Electrophoresis of isolated DNA and RNA in agarose gels.
Isolation of total protein from human tumor cells and physiological cells.
Separation of the proteins using electrophoretic methods - polyacrylamide gel electrophoresis SDS-PAGE.
Transfer of proteins and nucleic acids to nitrocellulose membranes. Techniques for Western-blot, Northern-blot, Southern-blot.
The use of genetically modified micro-organisms purpose of the production of proteins. Vectors for overexpression of proteins. Spectrofluorymetric analysis of expression of GFP in E. coli under varying culture conditions - testing the effect of the composition of different microbiological culture media of the intensity of expression of the reporter gene to test the strength of the promoter trc. Isolation of GFP and its spectrophotometric and electrophoretic analysis
Amplification of nucleic acids. The function of DNA and RNA polymerases. Restriction mapping and labeling of DNA. Genome sequencing. Molecular markers. Optimization of the gene construct for biotechnology: the selection of a vector, selection of a promoter, the choice of reporter gene / antibiotic resistance, selection of the appropriate host (HOST),., Cells, and organisms. Introduction of genetic material into the cells of pro- and eukaryotic cells. Cloning genes. Validation method for cloning genes (methods involving PCR). transfection of genes