Proteinbioreactor

Summary of the project

In our project we try to synthesize certain natural, modified and artificial proteins in a preparative scale. Prokaryotic and eukaryotic cells use very efficient reactions to produce a variety of different proteins in high yields. These reactions were used for the cell-free synthesis of proteins. The in vitro reaction is regulated by addition of a certain template (DNA or mRNA), which directs the synthesis of the desired protein only. The most important advantages of the cell-free proteinbiosynthesis in comparison to the in vivo expression are:

1. the distance from the gene to the final protein is very short,

2. the synthesis of toxic proteins is possible,

3. the site-directed isotope labeling allows to study their structure by NMR spectroscopy,

4. the site-specific incorporation of a non-natural or modified amino acid allows to investigate the function of the protein,

5. the purification of the products is less time consuming.

Selected publications

Stiege, W.; Erdmann, V.A.: The Potentials of the in vitro Protein Biosynthesis System. Journal of Biotechnology 41, 81-90 (1995).

Stiege, W.; Merk, H.; Gerrits, M.; Engelhard, M.; Kumagai, I.; Erdmann, V.A.: In vitro Protein Synthesis. Biol. Chemistry 377, 84 (1996).

Fuchs, U.; Stiege, W.; Erdmann, V.: Ribonucleolytic activities in the Escherichia coli in vitro translation system and in its separate components. FEBS Letters 414, 362-364 (1997).

Merk, H.; Stiege, W.; Tsumoto, K.; Kumagai, I.; Erdmann, V.A.: Cell-free expression of two single-chain monoclonal antibodies against lysozyme – Effect of domain arrangement on the expression. J. Biochem. 125, 328-333 (1999).

Last update: 29.1.2001