Part:BBa_K2644002

arsenic sensitive circuit

A circuit which is sensitive to As(III) and can express fluorescent protein smURFP

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
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 198
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Usage

Arsenic sensitive plasmid is a circuit which is sensitive to As(III) and express smURFP as a feedback so that we can indirectly educe As(III) concentration through detecting the fluorescent intensity of smURFP. It's composed of 4 small parts, including promoter J23104, RBS and ArsR gene(BBa_K2644010), promoter ArsR along with RBS(also named ArsR operator, BBa_K2644004), smURFP gene(BBa_K2644007) and HO-I gene(BBa_K2328062).

Biology

In order to detect trivalent arsenic ions(As(III))，we choose a kind of arsenic binding repressor protein--ArsR. ArsR binds the promoter in the absence of arsenite and acts as a repressor, and dissociates from the promoter in the presence of arsenite. Connecting with constitutive promoter J23104, the ArsR gene can work and bind the ArsR promoter in front of smURFP gene stably. When arsenite comes, the ArsR promoter will no longer be repressed and the smURFP+HO-I part will produce FPs which is mostly proportional to As(III) concentration. Fluorescence intensity of 660nm (ultra-red light) can be measured through the laser confocal microscopy.

Reference

[1] Davide Merulla1, Nina Buffi2, Siham Beggah1, Fre ́ de ́ ric Truffer3, Martial Geiser3, Philippe Renaud2 and Jan Roelof van der Meer1. Bioreporters and biosensors for arsenic detection. Biotechnological solutions for a world-wide pollution problem. [J].Current Opinion in Biotechnology 2013, 24:534–541 [2] John V Frangioni. In vivo near-infrared fluorescence imaging. [J].Current Opinion in Chemical Biology 2003, 7:626–634 [3] Rodriguez EA,Tran GN , Gross LA, et al. A far-red fluorescent protein evolved from a cyanobacterial phycobiliprotein .[J].NATURE METHODS,2016:763-769.