Learning objectives
The aim of the course is to provide an in-depth knowledge of the recombinant DNA technology.and his applications in bacteria, plant and animal field.
The course is divided in three parts:
1. The basics of recombinant DNA technology and main applications in bacteria
2. Application of recombinant DNA technology in plants
3. Application of recombinant DNA technology in animals
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
The course will provide in-depth knowledge on the methodologies and strategies underlying recombinant DNA technology.
The following topics will be covered during the course:
1. Basic aspects on the recombinant DNA technology:
- 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-based 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. Recombinant DNA technology in plants:
- Cloning vectors for plants
- Transformation methods for plants
- Transgenic plants
3. Recombinant DNA technology in animals:
- Cell cultures and selection systems for animal cells
- Gene transfer into animal cells
- Plasmid vectors for animal cells, amplicons and expression vectors
- Viral vectors and retroviral vectors for animal cells
- Gene therapy: principles and methods; experimental models and main experimental results.
Full programme
- - -
Bibliography
- Gene Cloning and DNA Analysis: An Introduction, 8th 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
The course is organized in:
1. theoretical classes by the teacher
2. seminars
3. training in laboratory
Assessment methods and criteria
Evaluation of the expected learning achievements will be based on:
1. a written exam on the basics of recombinant DNA technology and his application in bacteria;
2. an oral examination on the applications of recombinant DNA technology in plant and animal field.
To take the exam, the student must have undergone the laboratory training and prepare a "laboratory report" that will be evaluated during the examination.
The "laboratory report", the written and oral exams allow to verify the learning ability, independence of judgements, the acquisition of proper terminology, knowledge and application of knowledge to solve practical problems. The oral examination also verifies the communication skills.
Other information
- - -
