Bacteria as protein factories

Aim

The students will get hands-on experience of protein production using prokaryotic cell-based expression systems. The course presents methods to express genes, which have gene products that are known, unknown or not well characterized, and to analyze the gene products. Theories behind the methods are discussed and experiments are performed. The methods to be presented include: affinity purification of tagged proteins; generation and analysis of gene fusions; molecular cloning; polymerase chain reaction; metabolic engineering, seleno-methionine incorporation for MAD phasing or 13C, 15N-metabolic labeling for NMR studies; optimization of recombinant gene expression in Escherichia coli. Strategies for improving production of "difficult" proteins will be presented. Lectures by the instructors will cover the current status of the gene expression field, and theoretical aspects of the methodology. Guest lecturers will discuss contemporary problems in proteome research and the use of non-crystallographic methods to determine topology of membrane proteins. Study format: lectures (ca 15 %), student presentations (ca 5 %), and laboratory exercises (ca 80 %).

Location & Organization

Organizer

FLÄK - The Research School in Pharmaceutical Sciences

Course Director

Claes von Wachenfeldt

Location / venue

Lund University (Faculty of Science)

Timing & Workload

Duration 1 week
ECTS points 3
Frequency Annual
 

Examination yes

Criteria

Is the course taught in English? yes
Is documentation available? (book, syllabus)? yes
Is the course open for external researchers? yes

More Information

http://www.biol.lu.se/cellorgbiol/postgrad/courses/course_23.html