Difference between revisions of "Part:BBa K1025003"
 
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<partinfo>BBa_K1025003 short</partinfo>
 
<partinfo>BBa_K1025003 short</partinfo>
  
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iGEM mut part is a plasmid constructed by 2013 iGEM Tsinghua-E team which is used for genome level in vivo high-diversity library construction.
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Thu-E Mutation part is a plasmid constructed by 2013 iGEM Tsinghua-E team which is used for genome level in vivo high-diversity library construction.
  
 
===Usage and Biology===
 
===Usage and Biology===
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In this vector, highly error-prone dnaQ mutant, mutD1 was cloned downstream of araBAD promoter so that we can control the mutation intensity of the target genome by the concentration of araBAD promoter’s inducer, L-arabinose in a strict manner. And the GFP downstream mutD was to test whether the mutD can be translated.  
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In this vector, highly error-prone dnaQ mutant, mutD[1] was cloned downstream of araBAD promoter so that we can control the mutation intensity of the target genome by the concentration of araBAD promoter’s inducer, L-arabinose in a strict manner. And the GFP downstream mutD was to test whether the mutD can be translated.  
  
 
===Functional Parameters===
 
===Functional Parameters===
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We measured the mutation increase induced by our mut part with the protocol described in our note from. To be brief, by measuring the reversion of rifampinresistance caused by mutation in genome, we accurately determined the mutation rate increase to be as big as four times compared with negative control (either bacteria without mutD or without arabinose input). The data is shown below.
 
We measured the mutation increase induced by our mut part with the protocol described in our note from. To be brief, by measuring the reversion of rifampinresistance caused by mutation in genome, we accurately determined the mutation rate increase to be as big as four times compared with negative control (either bacteria without mutD or without arabinose input). The data is shown below.
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[[File:mutD-1.png|200px|thumb|left|alt text]]
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[[File:mutD-1.jpg|375px|thumb|left]]
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[[File:mutD-2.png|200px|thumb|left|alt text]]
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[[File:mutD-2.png|455px|thumb|left]]
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K1025006 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K1025003 SequenceAndFeatures</partinfo>
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===Reference===
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[1] Schaaper, R. M. MECHANISMS OF MUTAGENESIS IN THE ESCHERICHIA-COLI MUTATOR MUTD5 - ROLE OF DNA MISMATCH REPAIR. Proc. Natl. Acad. Sci. U. S. A.85, 8126-8130, doi:10.1073/pnas.85.21.8126 (1988).

Latest revision as of 16:34, 25 September 2013

Thu-E Mutation Part

Thu-E Mutation part is a plasmid constructed by 2013 iGEM Tsinghua-E team which is used for genome level in vivo high-diversity library construction.

Usage and Biology

In this vector, highly error-prone dnaQ mutant, mutD[1] was cloned downstream of araBAD promoter so that we can control the mutation intensity of the target genome by the concentration of araBAD promoter’s inducer, L-arabinose in a strict manner. And the GFP downstream mutD was to test whether the mutD can be translated.

Functional Parameters

We measured the mutation increase induced by our mut part with the protocol described in our note from. To be brief, by measuring the reversion of rifampinresistance caused by mutation in genome, we accurately determined the mutation rate increase to be as big as four times compared with negative control (either bacteria without mutD or without arabinose input). The data is shown below.

MutD-1.jpg
MutD-2.png



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 478
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1770
    Illegal AgeI site found at 73
    Illegal AgeI site found at 350
    Illegal AgeI site found at 839
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 55
    Illegal SapI.rc site found at 1061


Reference

[1] Schaaper, R. M. MECHANISMS OF MUTAGENESIS IN THE ESCHERICHIA-COLI MUTATOR MUTD5 - ROLE OF DNA MISMATCH REPAIR. Proc. Natl. Acad. Sci. U. S. A.85, 8126-8130, doi:10.1073/pnas.85.21.8126 (1988).