iGEM Bielefeld 2015


Cell-Free Protein Synthesis

E. coli

Why do we need E. coli in synthetic biology?

As a chassis, it has countless advantages: It growth fast, it is amenable for genetic changes, and it is probably the best characterized organism in microbiology.

plate with E. coli

But has it disadvantages?

E. colis used for research are genetically modified organisms. You are not allowed to use it outside the lab because it might have an impact on environment.

dead E. coli

How can we separate the advantages of E. coli from the biosafety concerns?

We use the best features from E. coli and get rid of the unnecessary ones! We disrupt the cells, thereby disabling the bacteria to reproduce, and we collect the cell extract with its molecular machinery to produce proteins in vitro, that is without living cells!

plate with E. coli

Transcription and translation without any organism? How is that possible?

By applying one of iGEMs central ideas: Combination! For Cell Free Protein Synthesis (CFPS) you need:

  • A DNA template which carries all information about your protein of interest
  • The protein synthesis machinery made up of proteins and ribosomes
  • Amino acids, energy resources and cofactors to fuel the machinery.

At a glance

  • We managed to produce high amounts of superfolder GFP in selfmade E. coli extract and optimized the system throughout the summer.
  • We successfully modelled the cell extract in simbiology.
  • Our extract contains vanishingly low amounts of living cells. However, after a final treatment of the extract, we did not observe even one living cell.
  • We tested the effect of different heavy metals on cell free protein synthesis.