Difference between revisions of "Part:BBa K1980012"
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<p>This box is present at the promoter regions of the copper exporting ATPase CopA, some molybdenum cofactor synthesis genes and the periplasmic copper oxidase protein CueO.<sup>(2)</sup></p>
 
<p>This box is present at the promoter regions of the copper exporting ATPase CopA, some molybdenum cofactor synthesis genes and the periplasmic copper oxidase protein CueO.<sup>(2)</sup></p>
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[[Image:Pmg_flow_cyto_oxford_sam_2016.jpeg|400px|thumb|centre|Schematic of the construct and how it works]]
  
 
==Experience==
 
==Experience==

Revision as of 21:46, 22 October 2016


pCopA MymT sfGFP with divergent CueR

This part is our chelator MymTsfGFP expressed from this copper-sensitive promoter pCopA with the regulator of pCopA (CueR) expressed divergent from the promotor.

Sequence and Features:


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 438
    Illegal NheI site found at 461
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1175
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Usage and Biology

E. coli cells use a protein called CueR to regulate the cytoplasmic copper concentration. CueR is a MerR-type regulator with an interesting mechanism of action whereby it can behave as a net activator or a net repressor under different copper concentrations through interaction with RNA polymerase(1). CueR forms dimers consisting of three functional domains (a DNA-binding, a dimerisation and a metal-binding domain). The DNA binding domains bind to DNA inverted repeats called CueR boxes with the sequence:

CCTTCCNNNNNNGGAAGG

This box is present at the promoter regions of the copper exporting ATPase CopA, some molybdenum cofactor synthesis genes and the periplasmic copper oxidase protein CueO.(2)


File:Pmg flow cyto oxford sam 2016.jpeg
Schematic of the construct and how it works

Experience

Microscopy performed on this part at different copper concentrations. Fluorescent, differential interference contrast channels and a composite shown



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

(2) Yamamoto K, Ishihama A. (2005) “Transcriptional response of Escherichia coli to external copper.” Mol Microbiol. 2005 Apr;56(1):215-27