Ateneo - UPO

Description and practical demonstration of techniques that are commonly used to analyze proteins contained in cell extracts, as well as recombinant proteins that have been over-expressed in heterologous systems and purified to homogeneity.

Scientific articles, review articles and material provided directly by the lecturer.

Enable students to become familiar with some of the protein analysis techniques widely used in research laboratories working in the fields of molecular and cellular biology, pharmacology, and biotechnology.

Knowledge of protein biochemistry, cell and molecular biology and methods for the design and production of recombinant DNA molecules.

The course is divided into three sections: lectures, exercises sessions and practical laboratory work. Audio-visual materials (in English) will be used during the lectures. Students will be trained in the use of publicly accessible databases, and encouraged to engage in group discussions to interpret the experimental results obtained, in a collaborative manner.

Students with physical disabilities, Learning Disabilities or Special Education Needs can request specific services and tools via the Staff Sviluppo e Coordinamento Carriere e Servizi alle Studentesse e agli Studenti, consulting the University webpage: https://www.uniupo.it/en/services/services-students-physical-or-learning-disabilities Students with disabilities, learning disabilities or special education needs, once they have contacted the University Staff, can refer to the tutor in charge of the course to define the examination modalities, concerning academic aspects.

WRITTEN EXAM in the form of open-ended questions relating to the theoretical topics covered, which will be included in the written exam for module FA0429 - molecular biology, starting from the summer exam session. Students who decide to take the partial exam for module FA0429 - Molecular Biology in the February exam session, may freeze their grade until the autumn session, and independently take the part of the exam relating to module FA0454. The accuracy, completeness and precision of the content and the drafting of the “laboratory notebook” will also be assessed.

Lectures - In-depth analysis of the use of biophysical techniques for the study of recombinant proteins and interactions between biomolecules: Differential Scannning Fluorimetry (DSF), with examples; Biolayer Interferometry (BLI), with examples. Practical exercises (in groups, in the classroom): design and drafting of an experimental protocol for the expression in recombinant form and the analysis of a protein of interest (chosen by the lecturer), based on the study and discussion of scientific articles reporting on the in vitro characterisation of functionally equivalent proteins. Lab activity - SDS-PAGE and western blot analysis of cell extract samples obtained from experiments conducted in module FA0455 - Applied biological techniques with laboratory. Purification, by IMAC chromatography, of a recombinant protein from clarified bacterial extracts.

At the end of the course, students will have acquired knowledge and understanding of the topics covered and developed the ability to independently apply this knowledge to the design of biochemical and functional protein analysis procedures (choosing the appropriate experimental conditions, instrumentation and methodology for conducting experiments) and will be able to write reports on the activities carried out.