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We characterized <a href="http://parts.igem.org/Part:BBa_K1333108">BBa_K1333108</a> as part of the genome wide mutators | We characterized <a href="http://parts.igem.org/Part:BBa_K1333108">BBa_K1333108</a> as part of the genome wide mutators | ||
<a href="http://parts.igem.org/Part:BBa_K2082116">BBa_K2082116</a> and <a href="http://parts.igem.org/Part:BBa_K2082117">BBa_K2082117</a>. | <a href="http://parts.igem.org/Part:BBa_K2082116">BBa_K2082116</a> and <a href="http://parts.igem.org/Part:BBa_K2082117">BBa_K2082117</a>. | ||
− | Our | + | Our characterization is based on <a href="https://2016.igem.org/Team:Bielefeld-CeBiTec/Results/Mutation/Reversion">reversion assays</a>, showing the mutation rate of dnaQ926. |
<figure class="figure"> | <figure class="figure"> | ||
<img src="https://static.igem.org/mediawiki/2016/3/3a/Bielefeld_CeBiTec_2016_10_19_mutation_reversion_global_rates_final.png" class="figure-img" /> | <img src="https://static.igem.org/mediawiki/2016/3/3a/Bielefeld_CeBiTec_2016_10_19_mutation_reversion_global_rates_final.png" class="figure-img" /> |
Revision as of 00:42, 20 October 2016
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DnaQ926 - BBa_K1333108
Introduction
DnaQ is part of the DNA polymerase III and responsible for the proofreading activity of this complex. The dnaQ926 variant
loses this activity through mutation of two function essential amino acids inside the active site. The complete loss of proofreading as well as the resulting saturation of mismatch-repair makes dnaQ926 the single strongest mutator gene known (Fijalkowska und Schaaper 1996) (see her for more informations)
is part of our genome wide mutator BBa_K2082117, but we also used it as a standalone mutator in BBa_K2082116. After initially designing our genome wide mutator we looked for mutagensis gene already in the iGEM partsreg and found several candidates. The most prominent one is iGEM SYSU-China 2014's dnaQ926 BBa_K1333108. Furthermore, we use iGEM Uchicago IGSB2014's emrR and dam in our large genome wide mutator.
is part of our genome wide mutator BBa_K2082117, but we also used it as a standalone mutator in BBa_K2082116. After initially designing our genome wide mutator we looked for mutagensis gene already in the iGEM partsreg and found several candidates. The most prominent one is iGEM SYSU-China 2014's dnaQ926 BBa_K1333108. Furthermore, we use iGEM Uchicago IGSB2014's emrR and dam in our large genome wide mutator.
Characterization
We characterized BBa_K1333108 as part of the genome wide mutators
BBa_K2082116 and BBa_K2082117.
Our characterization is based on reversion assays, showing the mutation rate of dnaQ926.
Furthermore, we determined precise dnaQ926 mutational spectrum via high-throughput sequencing.
In combination we added substentially to BBa_K1333108 characterization thus enabling future iGEM teams to use this part as an in vivo mutator.