Syllabus
Examples of experiments: 1) Determination of glycogen concentration in rat liver after starvation: the experimental aim is to isolate and determine glycogen content in rat liver. The glycogen content in the livers of two individuals, normally fed rats and short-term fasted rats, will be compared.
Glycogen will be separated from mono- and oligosaccharides and hydrolyzed in acidic conditions. The resulting glucose will be determined spectrophotometrically using an enzymatic test. 2) Isolation of recombinant protein, SDS-PAGE electrophoresis: the experimental aim is to isolate GST-fusion protein (14-3-3 or beta-galactosidase) from a bacterial culture of Escherichia coli.
Recombinant GST-fusion protein will be isolated from bacterial culture of E. coli by affinity chromatography. The quality of the individual purification steps will be verified by SDS-PAGE electrophoresis. 3) Phospholipids in biological membranes: the experimental aim is to compare the quantitatively different content of phospholipids in the membranes of erythrocytes of ruminant and non-ruminant animals.
Phospholipids will be divided into basic groups by thin layer chromatography and the relative proportion of selected phospholipid species will be determined quantitatively by densitometric image analysis.
Annotation
The course provides a working knowledge of biochemistry, focusing on practical aspects of the field. It covers basic biochemical techniques used in modern experimental biology, e.g. chromatography, spectrophotometry, and electrophoresis applied to the analyses of lipids, carbohydrates, proteins and enzymes in various biological materials.
Students are given the opportunity to perform simple biological experiments independently. General outline for each experiment: The introduction - is organized as interactive seminar to update the theoretical background and practice biochemical computations.
The experiment - is carried out by students independently (2 in a group). The analysis and presentation of data - written report is prepared by the students.
The discussion - interactive seminar to evaluate results and discuss problems. Examples of experiments: 1) Determination of glycogen levels in the mouse liver after starvation. 2) Isolation of recombinant protein, SDS-PAGE electrophoresis. 3) Quantitative analysis of erythrocytes phospholipids in ruminant and non-ruminant mammals.