Teaching code

4S02696

Credits

4

Coordinator

Not yet assigned

Language

Italian

Scientific Disciplinary Sector (SSD)

BIO/10 - BIOCHEMISTRY

Moodle Seminars 0

To show the organization of the course that includes this module, follow this link:  Course organization

The teaching is organized as follows:

Program

------------------------ UL: theory ----------------------- - Purification of proteins Ionic properties of amino acids and proteins. Isoelectric point. Sample preparation. Cell disruption methods and production of initial crude extracts. Protein solubilization methods. Saline swabs. Fractionation and precipitation techniques. Filtration, dialysis, sample concentration. Spectroscopic techniques. Properties of electromagnetic radiation. Light-matter interaction. States and processes involved in absorption, emission and decay phenomena. Ultraviolet and visible absorption spectroscopy. Qualitative and quantitative aspects of light absorption. Colorimetric and spectroscopic methods applied to the determination of protein concentration. Spectrophotometers. Emission and fluorescence spectroscopy. Intrinsic and extrinsic fluorophores. Fluorescent proteins (Green fluorescent proteins) Spectrofluorimeters. Fluorescence resonance energy transfer (FRET). Circular dichroism. Chromatographic techniques. Principles of chromatography. The chromatogram. Parameters that determine chromatographic performance. Column chromatography: ion exchange, molecular exclusion, affinity, hydrophobic interaction and their applications. Purification of fusion proteins with electrophoretic techniques. General principles and electrophoretic mobility. Support materials. Electrophoresis of nucleic acids. Protein electrophoresis. SDS PAGE. Electrophoresis under native conditions. Protein staining on gel. Protein blotting (western blotting). Notes on: Isoelectrofocusing. Capillary electrophoresis. Two-dimensional gel electrophoresis. ------------------------ UL: laboratory ----------------------- - 1) Preparation of buffers for the study of protein molecules 2) Colorimetric and spectroscopic methods for the determination of protein concentration. 3) Determination of the kinetic parameters of acid phosphatase. 4) Determination of the molar extinction coefficient of NADPH. 5) Electrophoretic methods: Polyacrylamide gel and SDS gel electrophoresis of proteins 6) WESTRN-BLOT ------------------------ UL: laboratory ---- -------------------- 1) Colorimetric and spectroscopic methods for the determination of protein concentration. 2) Determination of the kinetic parameters of acid phosphatase. 3) Determination of the molar extinction coefficient of NADPH. 4) Molecular exclusion chromatography to study the molecular weight of an unknown protein. 5) Electrophoretic methods: Polyacrylamide gel and SDS gel electrophoresis of proteins 6) WESTRN-BLOT

Didactic methods

------------------------ UL: theory ----------------------- - The course is structured in frontal theoretical lessons and guided exercises in the biochemistry laboratory. In particular, a total of 40 hours of teaching are foreseen (4 CFU) of which 16 hours of lessons and 24 hours of practical laboratory exercises. Lessons take place weekly in the classroom and the presentation takes place through the use of Power Point slides. For the laboratories, the students are divided into small groups. ------------------------ UL: laboratory ----------------------- - Practical laboratory experiences, with small groups of students ------------------------ UL: laboratory ------------ ------------ Small group laboratory experiences

Learning assessment procedures

------------------------ UL: theory ----------------------- - The final exam (written) will focus on all the topics of the program. The student must demonstrate that they understand and are able to use the fundamental concepts of each topic. ------------------------ UL: laboratory ----------------------- - The topics covered during the laboratories may be examined for the "Biochemistry and Analytical Biochemistry" course. ------------------------ UL: laboratory ----------------------- - The topics covered during the laboratories may be examined for the "Biochemistry and Analytical Biochemistry" course.

Evaluation criteria

------------------------ UL: theory ----------------------- - The written exam consists of 11 multiple choice and/or open questions, 3 points are assigned for each correct answer ------------------------ UL : laboratory ------------------------ The student's ability to perform basic calculations for the preparation of solutions used in the laboratory or for the calculation of dilutions. Furthermore, open or multiple choice questions will be asked to verify learning. ------------------------ UL: laboratory ----------------------- - The objective of the exam is to verify the level of knowledge and depth of the topics of the course program and the reasoning ability developed by the student.

Exam language

Italiano