Part:BBa_K2555002

pBAD-RBS-CueR

This construct contains pBAD (L-arabinose inducing promoter) with RBS and CueR (a transcription factor that binds onto pCopA of BBa_K2555000). E. coli cells use CueR to regulate the cytoplasmic copper concentration. Arabinose concentration controls the expression of CueR in the construct.CueR behaves as a net activator or a net repressor under different copper concentrations(1). BBa_K2555002 and BBa_K2555000 is used for construction of BBa_K2555004.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1311
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1250
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1085
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 1067
  

Method

For the part plate reader was done. A plate reader is able to take optical density (OD600nm) . OD is a measure of bacterial growth.

Results

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The results of a OD600 with different arabinose concentrations after five hours. Error bars show standard deviation of three repeats.

The OD600 value of the construct did not change significantly with the increase of Arabinose concentration. The increased expression of the CueR protein did not burden the bacteria, thereby affecting their growth.

References

(1)Danya J. Martell, Chandra P. Joshi, Ahmed Gaballa, Ace George Santiago, Tai-Yen Chen, Won Jung, John D. Helmann, and Peng Chen (2015) “Metalloregulator CueR biases RNA polymerase’s kinetic sampling of dead-end or open complex to repress or activate transcription” Proc Natl Acad Sci U S A. 2015 Nov 3; 112(44): 13467–13472.