Difference between revisions of "Part:BBa K389015"

 
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<partinfo>BBa_K389015 short</partinfo>
 
<partinfo>BBa_K389015 short</partinfo>
  
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This BioBrick is for testing the virA/G signaling system. The read-out is the luciferase <partinfo>K389004</partinfo>. This system contains an unmutated ''virA'', so it is possible to measure, how the natural virA/G system works and reacts.  
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This BioBrick contains a complete VirA/G receptor system with a firefly luciferase (from Promega's pGL4.10[luc2] vector) under the control of a ''vir'' promoter as reporter gene.  
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Input: acetosyringone
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Output: expression of [[Part:BBa_K389004 | luciferase]]
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<!-- Add more about the biology of this part here
 
 
===Usage and Biology===
 
===Usage and Biology===
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This BioBrick is for testing, characterizing and measuring the VirA/G signaling system. This system contains an unmutated ''virA'', so it is possible to measure how the natural VirA/G system works and reacts. It is also possible to determine the basal transcription of the ''vir'' promoter and its activity after inducing the system with acetosyringone.
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===Important parameters===
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<center>
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{|{{Table}}
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!Experiment
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!Characteristic
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!Value
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|-
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|rowspan="4"|[[Part:BBa_K389015:Experience#Transfer function | Transfer Function]]
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|Maximum induction level
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|2.2 fold
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|-
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|Maximum induction level reached
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|200 µM acetosyringone
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|-
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|Hill coefficient
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|1.09
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|-
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|[[Switch Point|Switch Point]]
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|31.6 µM acetosyringone
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|-
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|rowspan="3"|[[Part:BBa_K389015:Experience#Growth functions and Luciferase expression for BBa_K389015 | Doubling time / h]]
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|without plasmid
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|1.98
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|-
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|carrying K389015
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|2.24
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|-
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|carrying K389015 with 400 µM acetosyringone
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|2.67
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|-
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|rowspan="2"|Response time
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|Induction: [[Part:BBa_K389015:Experience#Response time | exponential phase]]
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|>1 h
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|-
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|Induction: [[Part:BBa_K389015:Experience#Data Analysis | begin of cultivation]]
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|max. induction at OD<sub>600</sub> = 1 +/- 0.5
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|-
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|rowspan="3"|[[Part:BBa_K389015:Experience#Plasmid conformation analysis | Conformation analysis]]
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|ratio ccc monomer / %
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|91
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|-
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|ratio ccc dimer / %
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|3.7
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|-
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|ratio oc forms / %
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|5.3
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|-
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|}
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</center>
  
 
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Latest revision as of 08:32, 27 October 2010

VirA/G reporter device luc

This BioBrick contains a complete VirA/G receptor system with a firefly luciferase (from Promega's pGL4.10[luc2] vector) under the control of a vir promoter as reporter gene.

Input: acetosyringone

Output: expression of luciferase


Usage and Biology

This BioBrick is for testing, characterizing and measuring the VirA/G signaling system. This system contains an unmutated virA, so it is possible to measure how the natural VirA/G system works and reacts. It is also possible to determine the basal transcription of the vir promoter and its activity after inducing the system with acetosyringone.


Important parameters

Experiment Characteristic Value
Transfer Function Maximum induction level 2.2 fold
Maximum induction level reached 200 µM acetosyringone
Hill coefficient 1.09
Switch Point 31.6 µM acetosyringone
Doubling time / h without plasmid 1.98
carrying K389015 2.24
carrying K389015 with 400 µM acetosyringone 2.67
Response time Induction: exponential phase >1 h
Induction: begin of cultivation max. induction at OD600 = 1 +/- 0.5
Conformation analysis ratio ccc monomer / % 91
ratio ccc dimer / % 3.7
ratio oc forms / % 5.3

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
    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