Learning outcomes of the course unit
KNOWLEDGE AND UNDERSTANDING
Students will gain theoretical and practical principles at the base of the main methodologies used for the study and analysis of proteins and nucleic acid.
APPLYING KNOWLEDGE AND UNDERSTANDING
At the end of the course, the students will have the necessary tools to address the complex molecular analysis of issues of interest in the field of food science.
Students will acquire the ability to evaluate and interpret experimental data and to develop a scientific approach to bioethical problems related to the world of food (eg. GMOs)
Students will be provided with the means to communicate experimental scientific results
ABILITY TO LEARN
Students will be provided with the tools to develop a scientific approach in carrying out experiments and interpreting and formalizing the results.
Students should have a solid background in Biochemistry
Course contents summary
The course will cover the main methods used in the study and analysis of proteins and nucleic acids and their application in food analyses.The course also includes classroom exercise activity.
The following topics are considered:
-Introduction to nucleic acid structure and stability.
-Methods of extraction and purification of nucleic acids.
-Electrophoresis of nucleic acids.
-Enzymes used for DNA manipulation: DNA polymerases, nucleases, ligases. Restriction enzymes and RFLP.
-DNA sequencing: chemical degradation method, chain termination method, pyrosequencing, thermal cycle sequencing, NGS sequencing.
-Qualitative and quantitative PCR (Polymerase chain reactions) and RT-PCR techniques.
-Fingerprinting techniques: RAPD, RFLP, AFLP.
-Cloning of exogenous genes, transformation of host organisms and selection of recombinants. Recombinant proteins production.
-Probe preparation and labelling for Southern e Northern Blotting and Microarray
-Methods used in the preparation of transgenic organisms and recombinant proteins for the agri-food sector. Bt corn and RoundUp ready soybean. Analytical methods for the determination of authorised and non-authorised GMOs in food products. Antisense technology. Cloned animals.
-Protein extraction and purification.
-Differentiated protein precipitation. Protein chromatographic separation techniques: hydrophobic interaction, ion exchange, dimensional exclusion and affinity/immunoaffinity chromatographies.
-Protein electrophoretic separation techniques: polyacrylamide gel electrophoresis, isoelectric focusing and two-dimensional electrophoresis.
-Immunochemical methodologies. Overview of the biochemical basis of immune response and of the structural/functional properties of antibodies. Preparation of polyclonal and monoclonal antibodies. Utilisation of antibodies as analytical reagents: immunoprecipitation, immunodiffusion, immunoelectrophoresis, Western Blotting, immunoenzymatic tests (ELISA).
-Biochemical basis of BSE and analyses methodology.
Lessons will be organized face-to-face with the possibility of using the lessons also remotely in asynchronous mode (uploaded on the Elly page of the course). The teaching will be carried out through lectures in the classroom with the help of slides that will represent teaching material, in addition to the recommended text. The slides will be available online on the website https://elly.saf.unipr.it/2020/ in pdf format for students.
Assessment methods and criteria
Written exam followed by possible oral exam.
The written exam, lasting 2 hours, includes 4 exercises, two questions for which a short and schematic answer is required and two questions for which more detailed answers are required.
The exercises and questions concern the fundamental concepts addressed in the course and have equal weight on the definition of the final grade.
The exam results are published on the ESSE3 portal (https://unipr.esse3.cineca.it/Home.do) within a reasonable time compatible with the number of students enrolled. Students will be able to view the written exam by making an appointment with the teacher. At the student's choice, it will be possible to integrate the written exam with an oral exam.
If it is impossible to take the written exam in classroom due to rules imposed by the University, examinations will be carried out remotely by using Teams platform. The assessment of the level of knowledge acquired also takes into consideration how the student is able to express himself correctly, with the specific scientific language of biochemistry.
In the event of health emergency, teaching and examination methods could undergo changes that will be promptly communicated on Elly platform and on the course website. Lecture records will be available until the end of the academic year.