Experiment and quantitative modeling of cell-free gene expression dynamics

Genexpression that is the cellular synthesis of proteins is
comprised of the sub-steps tran- scription (mRNA synthesis based on
the DNA master), translation (protein synthesis based on the mRNA)
and protein folding. Owing to the large number of interactions
between individual components this process is very complex in vivo
and therefore mathematical modeling is extremely laborious. By
means of simpli�ed in vitro model systems individual aspects of
cellular gene expression can be studied in detail. Cell-free gene
expression denotes the biochemical synthesis of proteins in vitro.
Cell-free systems are comprised of the predominant components of
the cellular transcription and translation machinery like
polymerases, ribosomes, amino acids and nucleotides dissolved in
bu�er. These systems are either cellular extracts or are
reconstituted from puri�ed components. In this thesis gene
expression in a cell-free system was used to develop a quantitative
model system of the gene expression kinetics in vitro. To this end
techniques were developed that enable reproducible quantitative
fuorescence measurements of mRNA and protein synthesized in a
cell-free system.