Lehrveranstaltung: Mutagenesis and Overexpression, Practical (1502-482)

Personen:

• Prof. Dr. Lutz Fischer (verantwortlich)
• Dr. Sabine Lutz-Wahl (begleitend)

Lehrform:

Praktikum

SWS:

5

Inhalt:

The students
- will learn strategies of cloning genes from target enzymes in suitable expression systems
- will learn methods for mutagenesis of genes
They will learn following methods: extraction and purification of genomic DNA and Plasmid-DNA, amplification of DNA by PCR, performance of site-specific mutagenesis by QuikChange PCR, error prone PCR, analytical and preparative agarose gel electrophoresis, restriction and ligation of DNA, transformation, DNA sequencing, cultivation of recombinant strains in shaking flasks, determination of enzyme activity and protein concentration, SDS polyacrylamide gel electrophoresis

Literatur:

Brown, T.A.: Gene cloning and DNA analysis, 6. Auflage, John Wiley and Sons, 2010
Glick, B. R., Pasternak, J.J.: Molecular Biotechnology - Principles and application of recombinant DNA, 3. Auflage, ASM, Washington, 2003
Sambrook, J., Fritsch, E. F., Maniatis, T.: Molecular Cloning, 3. Auflage, Cold Spring Harbor, New York, 2001
current scientific publications (will be provided)

Veranstaltungsort:

Hohenheim

Modul:

• 1502-480 Mutagenesis and Overexpression of Enzymes (Pflicht)