BIC-10306 Practical Biological Chemistry

Course

Credits 6.00

Language of instruction:

Dutch

Contents:

This introductory practical course provides basic knowledge and skills. Lectures provide the theoretical background of the applied techniques and the experiments to be performed. Subjects are: techniques of gene cloning, structure and function of nucleic acids and proteins, structure and expression of eukaryotic genes, electrophoresis, PCR, spectrophotometry and fluorescence. The practical consists of two parts: one molecular-biological and the other biochemical. The molecular-biological part is dedicated to: isolation and analysis of DNA, RNA and proteins, both from prokaryotes (E. coli) and eukaryotes (tomato); construction of a physical map of a plasmid by enzyme digestions and electrophoresis; detection techniques like PCR and the use of computer programs and internet in the analysis of structure and function of DNA (bio-informatics). The gene encoding one of the rubisco proteins will serve as a model in part of the experiments. The biochemical part comprises the following subjects: quantitative clinical-chemical enzyme assays (uric acid quantitation, enzymatic assay for a heart-attack), detection of iso-enzymes with iso-electric focussing (tissue analysis), following metabolic processes in vivo using fluorescence (regulation of glycolysis in yeast cells), a study of the reaction mechanism of enzymes with enzyme kinetics and computer graphics, design of an immunochemical hormone assay and design and execution of a fluorimetric enzyme activity assay.

Learning outcomes:

At the end of the course, students are expected to:
- have basic knowledge on genome structure and gene organization, transcription and RNA processing in eukaryotes;
- understand basic principles of DNA cloning, hybridization and PCR;
- have skills in basic techniques in molecular biology, such as DNA and RNA isolation, PCR, restriction enzyme digestions and analysis, agarose gel electrophoresis;
- have basic knowledge of DNA- and protein analysis software and perform simple database searches on the internet;
- understand the principles of spectrophotometry, fluorometry, immunological methods and electrophoresis and the use of these techniques in biochemical analyses;
- design and perform enzymatic assays;
- be able to calculate and interpret independently the results of their experiments.

Activities:

Attending lectures and participation to the practical course. During the practical course, experimental results are elaborated and a report is written wich will be mailed by the assistent. Students receive tasks that should be solved during the course by Internet searches.

Examination:

The final mark is based on the results of an exam, laboratory skills and on the quality of report of the practical part.

Literature:

A manual for the practical course Biological Chemistry is available at the secretary of the Laboratories for Biochemistry and Molecular Biology. The theoretical background of this course can be found in:
- Berg, Tymoczko & Stryer: 'Biochemistry' (6th ed, Kniphorst).