BIC-10306 Practical Biological Chemistry

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Studiepunten 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 (detection of tumours, 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 perform a fluorimetric enzyme activity assay.

Aims:

Obtaining basic knowledge on genome structure and gene organisation, transcription and RNA processing in eukaryotes; understanding basic principles of DNA cloning, hybridisation, and PCR; obtaining skills in basic molecular biological techniques such as DNA and RNA isolation and analysis and basic knowledge on standard DNA and protein analysis software. Learning biochemical techniques, as well as the theoretical background. At the end of this course, students are expected to handle the applied apparatuses independently, to perform analyses and to calculate and interpret the obtained results independently. Students are also expected to perform simple (database) searches on the Internet.

Activities:

Attending lectures and participation to the practical course. During the practical course, experimental results are elaborated and a (short) report is discussed next day (or as soon as possible) with the assistant. Students receive tasks that should be solved during the course by Internet searches.

Examination:

The practical course is marked according to the obtained practical results and the written reports (2/3) and by the result of a test at the end of the course (1/3). A minimal mark 5.5 is required for the test.

Literature:

A manual for the practical course Biological Chemistry is available at the secretary of the Laboratory for Biochemistry. The theoretical background of this course can be found in two textbooks: - Berg, Tymoczko & Stryer: 'Biochemistry' (5th ed, Kniphorst;
- Lodish et al: 'Molecular Cell Biology' (5th ed, Kniphorst).