Heterologous expression of proteins in bacteria. The laboratory practical applications are devoted to the production of two 15N labelled proteins which will be used to study their interaction through biophysics and biochemistry techniques such as CD, NMR and SEC. It will be presented some of the main docking programs, with particular reference to computational approaches based on the use of experimental biochemical information
No specific textbooks are available.
Book chapters and review articles on specific subjects will be provided to the students.
Learning Objectives
The course is aimed to provide the students with a basic knowledge of the molecular biology techniques for the protein expression in bacterial system. Techniques for the isotopic labelling of proteins. Cell-free. Knowledge of the techniques for structural characterization of protein-protein, protein-small molecule and protein-metal ion interactions. Analysis of UV-vis, circular dichroism and basic NMR spectra. Knowledge of the main computational programs for obtaining a structural model of a protein-protein or protein-small molecule complexes.
Prerequisites
Courses required: none
Courses recommended: none
Teaching Methods
Power point presentations of the lectures
Further information
Attendance at the laboratory is compulsory
Type of Assessment
Oral exam. The date of the exams can be found at the following web site
https://sol.unifi.it/docprenot/docprenot
Course program
Heterologous expression of proteins in E. coli. Protein overexpression into an E coli expression vector. Strain of E. coli, promotors, fusion proteins. Cell-free techniques. Production of membrane proteins and optimization of biomolecules for the production of vaccines. Isotope enrichment.
The laboratory practical applications are devoted to the production of two proteins which will be used to study their interaction through biophysics and biochemistry techniques such as CD, NMR and SEC.
It will be presented some of the main docking programs, with particular reference to computational approaches based on the use of experimental biochemical information