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| GENE0001-3 | Genetic engineering
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| Duration : | 30h Th, 20h Pr |
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| Credits/ECTS : |
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| Holder(s) : | Jacques Dommes, Joseph Martial |
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| Language : | French language |
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| Course contents : | 1. Introduction (J. Martial)
- Definitions
- Molecular biology
2. Manipulation of DNA fragments (J. Martial)
- Isolating DNA fragments
- Modification of DNA fragments
- Ligation of DNA fragments
- Identification and characteristics of DNA fragments
3. Genetic engineering of Prokaryotes (J. Martial)
- Cloning vectors for Escherichia coli
- Hybrid vectors
- Expression of prokaryotic genes
- Expression vectors for the production of heterologous proteins in E. coli
- Cloning and expression in Gram+ bacteria
4. Construction and screening of DNA libraries (J. Dommes)
- Number of clones in a DNA library
- Construction of a DNA library
- Identification (selection by complementation, selection markers)
- Screening of a DNA library by hybridisation
- Screening of a DNA library through the detection of a protein
5. Site-directed mutagenesis (J. Dommes)
- Random mutagenesis
- Site-directed mutagenesis using synthetic oligonucleotides
6. Specific application of PCR (J. Dommes)
- PCR
- Diagnostic applications
- DNA sequencing
- Modification of DNA fragments
- Analysis of point mutations
- Inverse PCR
- RT-PCR
- Quantitative PCR
7. Genetic engineering of yeasts and filamentous fungi (J. Dommes)
- Transformation methods in yeasts
- Vectors for Saccharomyces cerevisiae
- Vectors for Kluyveromyces
- Methylotrophic yeasts (Pichia, Hansenula)
- Protein secretion in yeasts
- Most frequent expression problems in yeasts
- Filamentous fungi
8. Plant genetic engineering (J. Dommes)
- Transformation via Agrobacterium tumefasciens and the Ti plasmid
- Direct transformation methods
- Phytovirus vectors
- Inverse genetics in plants
- Metabolic engineering
- « Protein pharming »
- Production of recombinant protein in unicellular green algae
9. Production of recombinant proteins in insect cells (J. Dommes)
- Promoters
- Vectors
- Expression of recombinant proteins in silkworms
10. Cloning in mammalian cells (J. Martial)
- Selection markers
- Transfection methods
- SV40 vectors
- Vaccinia vectors
- Alphavirus vectors
- Gene therapy
- Adenovirus vectors
- Herpes vectors
11. Transgenic animals (J. Martial)
- Transfection methods
- Usefulness of transgenic animals
- « Dolly » : Multiplication of one individual through transfer of a somatic nucleus in an enucleated ovule |
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| Course objective : | · To get acquainted with the main methods used in genetic engineering
· To get informed of the potential of genetic engineering in the study of living organisms, as well as in the improvement of species for human use and for the production of recombinant proteins. |
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| Prerequisites : | Molecular biology and genetics (courses of the bachelier in Biology). |
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| Workshops : | Practicals for this course are integrated into "Training periods or integrated practicals (including seminars)"(SSTG0009-1). |
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| Organization : | Lectures during first period. |
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| Written notes : | Powerpoint presentations and lecture texts (partial coverage of the course) available through the WebCT system (part of Prof. J. Dommes).
Documents also available for the part given by Prof. J. Martial. |
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| Assessment : | Written exam (first exam exam session, as well as second one). |
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| Contacts : | - Prof. J. Martial (jmartial@ulg.ac.be)
- Prof. J. Dommes (j.dommes@ulg.ac.be) |
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| Items online : |
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| Materials available on WebCT |
| Syllabus and Powerpoint slides (part of Prof. J. Dommes) |
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