Difference between revisions of "Part:BBa K389011"
 
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This part contains:  
 
This part contains:  
  
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- a mutated ''virG'' response regulator from the virA/G receptor system from ''Agrobacterium tumefaciens'' which works in ''Escherichia coli'' without an ''rpoA'' gene from ''A. tumefaciens''
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* a mutated ''virG'' response regulator from the VirA/G receptor system from ''Agrobacterium tumefaciens'' which works in ''Escherichia coli'' without an ''rpoA'' gene from ''A. tumefaciens''
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* a kanamycin resistance under the control of a ''vir'' promoter
  
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- a kanamycin resistance under the control of a ''vir'' promotor
 
  
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This device is used to screen ''virA'' receptor mutants. The ''virA'' receptor is mutated in the BBa_K389010 BioBrick. The better the receptor detects a substance, the more ''virG'' is phosphorylated and the stronger the kanamycin resistance from this BioBrick is expressed. The BioBricks BBa_K389010 and BBa_K389011 have to be used together in one cell.
 
  
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===Usage and Biology===
 
===Usage and Biology===
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This device is used to screen ''virA'' receptor mutants. The ''virA'' receptor gene is mutated in the <partinfo>BBa_K389010</partinfo> BioBrick. The better the receptor detects a substance, the more VirG is phosphorylated and the stronger the kanamycin resistance from this BioBrick is expressed. The screening system for a mutated ''virA'' gene contains this BioBrick in a plasmid with R6K ori and the BioBrick <partinfo>BBa_K389010</partinfo> in a plasmid with ColE1 ori. Both plasmids are transformed to and screened in ''E. coli'' EC100D because 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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Latest revision as of 12:40, 9 October 2010

VirA screening device

This part contains:

  • a mutated virG response regulator from the VirA/G receptor system from Agrobacterium tumefaciens which works in Escherichia coli without an rpoA gene from A. tumefaciens
  • a kanamycin resistance under the control of a vir promoter


Usage and Biology

This device is used to screen virA receptor mutants. The virA receptor gene is mutated in the BBa_K389010 BioBrick. The better the receptor detects a substance, the more VirG is phosphorylated and the stronger the kanamycin resistance from this BioBrick is expressed. The screening system for a mutated virA gene contains this BioBrick in a plasmid with R6K ori and the BioBrick BBa_K389010 in a plasmid with ColE1 ori. Both plasmids are transformed to and screened in E. coli EC100D because 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
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]