Learning objectives
The aim of the course is to illustrate the basic methods of recombinant technologies and their applications in microbes, plants and animals.
The course is divided in four parts:
1. Aims and importance of gene cloning. Methods and tools used in prokaryotic cells
2. Application of recombinant technologies in plants
3. Application of recombinant technologies in animals and for health
Prerequisites
Some previous knowledge of biology, botany, microbiology, biochemistry and chemistry is highly advisable, as well a suitable level of English to consult literature.
Course unit content
1. Basic module
Fundamental principles of gene cloning - Enzymes used for recombinant DNA technology - Isolation and purification of genomic and plasmid DNA -
Electrophoresis of nucleic acids - Cloning vectors for bacteria - Phage vectors: lambda and M13 - In vitro DNA synthesis (PCR) - Mutagenesis mediated by PCR - Eukaryotic cloning vectors - Construction and analysis of genomic libraries - mRNA purification and cDNA synthesis - Construction and analysis of cDNA libraries - DNA sequencing.
2. Cloning in plants
Cloning vectors for plant organisms – Transformation methods for plant organisms - Obtainment of transgenic plants.
3. Cloning in animal cells and applications in health field
Growth and properties of animal cells in culture - Selection systems for animal cells - Gene transfer into animal cells - Plasmid vectors for animal cells, amplicons and expression vectors - The fate of transferred DNA - Viral vectors and retroviral vectors for animal cells. Gene therapy: principles and methods - Experimental models – Some experimental results - Potential human models.
Full programme
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Bibliography
- Gene Cloning and DNA Analysis: An Introduction, 7th Edition. Terry A. Brown. Zanichelli
- Dai geni ai genomi. Dale, von Schantz. EdiSES
- Analisi dei geni e genomi. Richard J. Reece. EdiSES
- Ingegneria genetica. Principi e tecniche. S. Primrose et al. Zanichelli
- DNA Ricombinante - J.D. Watson et al. Zanichelli
- Molecular biotechnology: principles and applications of recombinant DNA. Bernard R. Glick
- Molecular biology: principles of genome function. Nancy L. Craig. Oxford university press
- Ingegneria Genetica. S.M. Kingsman, AJ Kingsman. Piccin
- Genetica generale e umana (volume secondo). L. De Carli, E. Boncinelli, G.A. Danieli, L. Larizza. Piccin
Teaching methods
It is conducted through:
1. theoretical classes by the teacher, followed by the individual work by the student for preparation
2. seminars
3. training in laboratory
4. individual work for preparation of examination
Assessment methods and criteria
Students can choose between two ways of examination:
1. a written exam on the whole programme;
2. the oral examination (Erasmus students; students who want to improve already existing grades).
Moreover, in both cases to be admitted to the oral examination the student should prepare a report on laboratory training that will be evaluated during the examination.
The written exam and the laboratory report verifies knowledge and application of knowledge. The oral examination verifies the knowledge acquired by the student and the communication skills. In all cases, the learning ability is verified.
Other information
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