Difference between revisions of "Part:BBa K389015:Design"

(Design Notes)
 
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===Design Notes===
 
===Design Notes===
 
* The ''virA'' gene is under the control a medium strong constitutive promoter so there is enough VirA receptor expressed but not too much so the cell suffers from the expression (VirA is a membrane protein)
 
* The ''virA'' gene is under the control a medium strong constitutive promoter so there is enough VirA receptor expressed but not too much so the cell suffers from the expression (VirA is a membrane protein)
 +
* The ''virG'' gene is mutated so it works without an ''rpoA'' subunit from ''Agrobacterium tumefaciens'' (YC Jung ''et al.'', 2004)
 
* double terminator (forward) to keep expression of luciferase gene by the constitutive promoter low  
 
* double terminator (forward) to keep expression of luciferase gene by the constitutive promoter low  
 
** double terminator <partinfo>BBa_B0017</partinfo> instead of <partinfo>BBa_B0015</partinfo> because of problems mentioned with <partinfo>BBa_B0012</partinfo>
 
** double terminator <partinfo>BBa_B0017</partinfo> instead of <partinfo>BBa_B0015</partinfo> because of problems mentioned with <partinfo>BBa_B0012</partinfo>
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===Source===
 
===Source===
  
''A. tumefaciens'' C58 TI-plasmid, parts.igem, Mr. Gene, Promega's pGL4.10luc2 plasmid
+
* ''virA'' from ''A. tumefaciens'' C58 (<partinfo>K389001</partinfo>)
 +
* ''virG'' gene synthesized by Mr. Gene (<partinfo>BBa_K389002</partinfo>)
 +
* ''vir'' promoter from ''A. tumefaciens'' C58 (<partinfo>BBa_K389003</partinfo>)
 +
* luciferase gene from Promega's pGL4.10[luc2] vector (<partinfo>K389004</partinfo>)
 +
* constitutive promoter, RBS and double terminator from parts.igem (<partinfo>BBa_J23110</partinfo>, <partinfo>B0034</partinfo>, <partinfo>BBa_B0017</partinfo>)
  
 
===References===
 
===References===
 +
YC Jung ''et al.'' (2004) Mutants of ''Agrobacterium tumefaciens virG'' Gene That Activate Transcription of ''vir'' Promoter in ''Escherichia coli'', ''Current Microbiol'' 49:334-340.

Latest revision as of 14:37, 17 October 2010

VirA/G reporter device luc


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
    Illegal NheI site found at 2581
    Illegal NheI site found at 2604
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1632
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 3769
    Illegal NgoMIV site found at 5113
    Illegal NgoMIV site found at 5134
    Illegal AgeI site found at 4837
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1768
    Illegal SapI.rc site found at 5019


Design Notes

  • The virA gene is under the control a medium strong constitutive promoter so there is enough VirA receptor expressed but not too much so the cell suffers from the expression (VirA is a membrane protein)
  • The virG gene is mutated so it works without an rpoA subunit from Agrobacterium tumefaciens (YC Jung et al., 2004)
  • double terminator (forward) to keep expression of luciferase gene by the constitutive promoter low
  • luciferase gene to show the activity of the vir promoter
  • this BioBrick is for measuring the behaviour of the natural VirA/G system

Source

References

YC Jung et al. (2004) Mutants of Agrobacterium tumefaciens virG Gene That Activate Transcription of vir Promoter in Escherichia coli, Current Microbiol 49:334-340.