Difference between revisions of "Part:BBa K389010"
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__NOTOC__
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<partinfo>BBa_K389010 short</partinfo>
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This BioBrick contains a VirA receptor from ''Agrobacterium tumefaciens'' C58 under the control of a medium strong constitutive promoter.  
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===Usage and Biology===
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We used this BioBrick to mutate ''virA'' by error-prone PCR so the mutated VirA receptor is expressed and ready to be screened for several substances.  
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The screening system for a mutated ''virA'' gene contains this BioBrick in a plasmid with ColE1 ori and the BioBrick <partinfo>K389011</partinfo> in a plasmid with R6K ori. Both plasmids will be transformed to and screened in ''E. coli'' EC100D. Plasmids with R6K ori are only replicated in pir+ or pir116 ''E. coli'' strains. Once a receptor with high sensitivity for a screened substance is found, the plasmids are isolated and transformed to ''e.g. E. coli'' TOP10. Because the R6K ori does not work in this strain because it is not a pir+ / pir116 strain, it is easy to separate the mutated ''virA'' BioBrick from the screening plasmid.  
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<!-- -->
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K389010 SequenceAndFeatures</partinfo>
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<!-- Uncomment this to enable Functional Parameter display
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===Functional Parameters===
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<partinfo>BBa_K389010 parameters</partinfo>
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<!-- -->

Latest revision as of 16:17, 10 May 2013

VirA receptor (expressed)

This BioBrick contains a VirA receptor from Agrobacterium tumefaciens C58 under the control of a medium strong constitutive promoter.


Usage and Biology

We used this BioBrick to mutate virA by error-prone PCR so the mutated VirA receptor is expressed and ready to be screened for several substances.

The screening system for a mutated virA gene contains this BioBrick in a plasmid with ColE1 ori and the BioBrick BBa_K389011 in a plasmid with R6K ori. Both plasmids will be transformed to and screened in E. coli EC100D. Plasmids with R6K ori are only replicated in pir+ or pir116 E. coli strains. Once a receptor with high sensitivity for a screened substance is found, the plasmids are isolated and transformed to e.g. E. coli TOP10. Because the R6K ori does not work in this strain because it is not a pir+ / pir116 strain, it is easy to separate the mutated virA BioBrick from the screening plasmid.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 647
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1632
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1768