DnaQ is part of the DNA polymerase III and is responsible for the proofreading activity of this complex. The dnaQ926 variant
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DnaQ is part of the DNA polymerase III and responsible for the proofreading activity of this complex. The <i>dnaQ926</i> variant
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loses this activity through mutation of two function essential amino acids inside the active site. The complete loss of proofreading
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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 <i>dnaQ926</i> the single strongest mutator gene known (Fijalkowska und Schaaper 1996) (<a href="">see her for more informations</a>)
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as well as the resulting saturation of mismatch-repair makes dnaQ926 the single strongest mutator gene known. (Fijalkowska und Schaaper 1996) (<a href="">see her for more informations</a>)
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<br>
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dnaQ926 is part of our genome wide mutator BBa_K2082117, but we also used it as a standaloone mutator in part BBa_K2082116. After initially designing our genome wide mutator we looked for
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</i> is part of our genome wide mutator BBa_K2082117, but we also used it as a standalone mutator in BBa_K2082116. After initially
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mutagensis gene already in the iGEM partsreg and found several ones. The most prominent one is <a href="https://2014.igem.org/Team:SYSU-China">iGEM SYSU-China 2014's</a> dnaQ926 <a href="http://parts.igem.org/Part:BBa_K1333108">BBa_K1333108</a>.
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designing our genome wide mutator we looked for mutagensis gene already in the iGEM partsreg and found several candidates.
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Furthermore we use <a href="https://2014.igem.org/Team:UChicago">iGEM Uchicago IGSB2014's</a> <a href="http://parts.igem.org/Part:BBa_K1500995">emrR</a> and <a href="http://parts.igem.org/Part:BBa_K1500994">dam</a> in our large genome wide mutator.
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The most prominent one is <a href="https://2014.igem.org/Team:SYSU-China">iGEM SYSU-China 2014's</a> dnaQ926 <a href="http://parts.igem.org/Part:BBa_K1333108">BBa_K1333108</a>.
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Furthermore, we use <a href="https://2014.igem.org/Team:UChicago">iGEM Uchicago IGSB2014's</a> <a href="http://parts.igem.org/Part:BBa_K1500995"><i>emrR</i></a> and <a href="http://parts.igem.org/Part:BBa_K1500994"><i>dam</i></a> in our large genome wide mutator.
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>.
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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
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Part of our charactization were <a href="https://2016.igem.org/Team:Bielefeld-CeBiTec/Results/Mutation/Reversion">reversion assays</a>, showing the mutation rate of dnaQ926.
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<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>.
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Our charactization based on <a href="https://2016.igem.org/Team:Bielefeld-CeBiTec/Results/Mutation/Reversion">reversion assays</a>, showing the mutation rate of dnaQ926.
<b>Figure 1: Mutagensis rate of <i>dnaQ926</i> (BBa_K2082116) and M6 (BBa_K2082117).</b>
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<b>Figure 1: Mutagensis rate of <i>dnaQ926</i> (<a href="http://parts.igem.org/Part:BBa_K2082116">BBa_K2082116</a>) and M6 (<a href="http://parts.igem.org/Part:BBa_K2082117">BBa_K2082117</a>).</b>
</figcaption>
</figcaption>
</figure>
</figure>
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Furthermore we determined dnaQ926 mutational spectrum via <a href="https://2016.igem.org/Team:Bielefeld-CeBiTec/Results/Mutation/Sequencing">high-throughput sequencing</a>.
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Furthermore, we determined precise <i>dnaQ926</i> mutational spectrum via <a href="https://2016.igem.org/Team:Bielefeld-CeBiTec/Results/Mutation/Sequencing">high-throughput sequencing</a>.
<b>Figure 2: Mutagenic spectrum of the genome wide mutators <i>dnaQ926</i> (BBa_K2082116) and M6 (BBa_K2082117).</b>
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<b>Figure 2: Mutagenic spectrum of the genome wide mutators dnaQ926 (<a href="http://parts.igem.org/Part:BBa_K2082116">BBa_K2082116</a>) and M6 (<a href="http://parts.igem.org/Part:BBa_K2082117">BBa_K2082117</a>). </b>
</figcaption>
</figcaption>
</figure>
</figure>
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In combination we added substentially to <a href="http://parts.igem.org/Part:BBa_K1333108">BBa_K1333108</a> characterization thus enabling coming iGEM Teams to use this part as an <i>in vivo</i> mutator.
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In combination we added substentially to <a href="http://parts.igem.org/Part:BBa_K1333108">BBa_K1333108</a> characterization thus enabling future iGEM teams to use this part as an <i>in vivo</i> mutator.
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'semrR 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 charactization based on reversion assays, showing the mutation rate of dnaQ926.
Figure 1: Mutagensis rate of dnaQ926 (BBa_K2082116) and M6 (BBa_K2082117).
Furthermore, we determined precise dnaQ926 mutational spectrum via high-throughput sequencing.
Figure 2: Mutagenic spectrum of the genome wide mutators dnaQ926 (BBa_K2082116) and M6 (BBa_K2082117).
In combination we added substentially to BBa_K1333108 characterization thus enabling future iGEM teams to use this part as an in vivo mutator.
