In the Evobody generating process we want to obtain a high affinity binding protein by directed evolution.
To further evolve binding proteins with an innate ability to interact with our target we want to mutagenize
them and thereby enable an ongoing adaptation to the target.
We successfully cloned two different approaches to in vivo mutagenesis, demonstrated and quantified
mutagenesis by reversion assays and determined the mutation spectrum by next-generation sequencing.
Cloning
We added several in vivo mutagensis parts to the iGEM partsreg
From our mutators we determined the mutation rate by reversion assays using a stop-beta-lactamase.
Figure 1: Mutagenesis rate of our mutagenesis systems as determined by beta lactamase reversion assays.
Furthermore we determined the mutation rate of our error prone polymerase I as well as from our genome wide mutator
BioBrick BBa_K2082117 by high-throughput sequencing.
Figure 2: Mutagenesis rate of our our error prone polymerase I and our genome wide mutator BBa_K2082117Figure 3: Mutagenic spectrum of our error prone polymerase I and our genome wide mutatorsBBa_K2082117 and BBa_K2082116
