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By adding <a href="http://parts.igem.org/wiki/Part:BBa_K2082106">BBa_K2082106</a> to the iGEM parstreg and characterizing it we hope | By adding <a href="http://parts.igem.org/wiki/Part:BBa_K2082106">BBa_K2082106</a> to the iGEM parstreg and characterizing it we hope | ||
− | to give coming iGEM teams the possibility to easily optimize their proteins by means of directed evolution | + | to give coming iGEM teams the possibility to easily optimize their proteins by means of directed evolution <i>in vivo</i>. |
</div> | </div> | ||
<div class="container text_header"><h4>Characterization</h4></div> | <div class="container text_header"><h4>Characterization</h4></div> |
Revision as of 10:55, 19 October 2016
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Error Prone Polymerase I - BBa_K2082106
Integral to our project is the directed evolution of our Evobodies. To continuously introduce new Evobody variants with potential
binding capabilities into the population we uses the in vivo mutagenesis capabilities of the error-prone polymerase I
BBa_K2082106.
This enzymes replace the normal polymerase I under certain conditions (further information) and introduces a high amount of mutations inside our Evobody coding sequence. In that the mutations are more frequently inside our Evobody sequence and rarely inside other genomic proteins this in vivo mutagenesis approach circumvents the main problems restraining in vivo mutagenesis: the unintentional mutagenesis of essential proteins of a bacteria.
By adding BBa_K2082106 to the iGEM parstreg and characterizing it we hope to give coming iGEM teams the possibility to easily optimize their proteins by means of directed evolution in vivo.
This enzymes replace the normal polymerase I under certain conditions (further information) and introduces a high amount of mutations inside our Evobody coding sequence. In that the mutations are more frequently inside our Evobody sequence and rarely inside other genomic proteins this in vivo mutagenesis approach circumvents the main problems restraining in vivo mutagenesis: the unintentional mutagenesis of essential proteins of a bacteria.
By adding BBa_K2082106 to the iGEM parstreg and characterizing it we hope to give coming iGEM teams the possibility to easily optimize their proteins by means of directed evolution in vivo.
Characterization
We characterized the error prone polymerase I per performing reversion assays, thus quantifiying the mutation rate.
Furthermore we analyzed error-prone polymerase I mutation spectrum using Illumina sequencing.
In combination we benchmarked the error-prone polymerase I by working out all necessary information to use this BioBrick in further directed evolution applications.