Part:BBa_K389015

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