Syllabus
1. Isolation of a protein - enzyme, determination of specific activity
2. Gel chromatography of proteins (separation, relative molecular mass determination)
3. SDS - polyacrylamide electrophoresis (determination of relative molecular mass of proteins)
4. Practical enzymology - kinetic characteristics, Michaelis constants, maximal reaction rate, pH optimum, activation and inhibition of enzymes, electrophoretic separation of isoenzymes and detection of the enzyme activity in gel
5. DNA manipulation (restriction analysis)
6. Oral presentation of one experiment
Annotation
Practical course in biochemistry is provided by Helena Ryšlavá, Daniel Kavan, Veronika Hýsková, Petr Novák, Petr Pompach, Josef Chmelík, Renata Ptáčková, Michal Rosůlek, Lukáš Slavata, Petra Darebná a Kateřina Bělonožníková
The students are dealing with basic biochemical methods used for protein and nucleic acid isolation and their characterization (extraction, centrifugation, precipitation, dialysis, protein content determination). Gel filtration is used for protein purification from a mixture or for relative molecular mass determination. The students determine the relative molecular mass of proteins by SDS polyacrylamide gel electrophoresis. Important part of the practical course is dealing with enzymes, determination of Michaelis constant, maximal reaction rate, pH optimum, activation and inhibition, electrophoretic separation of isoenzymes and their detection in gel.