Designed by: Kevin Yang Group: iGEM14_UChicago (2014-10-17)
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how you used this part and how it worked out.
dam is part of our genome wide mutator BBa_K2082117.
Characterization
We characterized BBa_K1500994 as part of the genome wide mutator
BBa_K2082116.
Our characterization is based on reversion assays.
Figure 1: Mutagensis rate of dnaQ926 (BBa_K2082116) and M6 (BBa_K2082117). The mutation rate of standalone dnaQ926 is (2.3±1.12)×10-6 bp-1×generation-1 induced and (6.67±4.09)×10-8 bp-1×generation-1 repressed. The mutation rate of M6 (assembly of six mutator genes amongst other dnaQ926) is (7.16±4.31)×10-6 bp-1×generation-1 induced and (2.51±3.18)×10-8 bp-1×generation-1 when repressed.
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). Teh mutagenic spectrum was determined by distribution all oberserved mutations to the different mutations (reference base &arr; mutated base). The tables show the mutation distribution in percent. n is the number of counted mutation events.
Other important information for everyone using BBa_K2082116 is that the construct decreases cell viability. Furthermore, even very low amounts of expression create a strong mutator phenotype thus use of a very tightly controlled promoter is recommended. For this application iGEM Bielefeld 2016 used a modified PBAD, further repressed by addition of 20 mM glucose.
