Difference between revisions of "Part:BBa M50440"

 
 
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<partinfo>BBa_M50440 short</partinfo>
 
<partinfo>BBa_M50440 short</partinfo>
  
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This is a genetically encodable pH probe that will report cystolic pH in vivo of mammalian, yeast, or E. coli cells, utilizing Bioluminescence Resonance Energy Transfer (BRET). The fluorophore protein luminesces ratiometrically dependent on the pH in the cell.
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This is a genetically encodable probe that will report cytosolic pH in vivo of mammalian, yeast, or bacterial cells, through Bioluminescence Resonance Energy Transfer (BRET) data. The device utilizes a Rluc8 luciferase-cpVenus fluorophore fusion, where the Rluc8 luciferase protein exhibits pH-dependent activity. The luciferase cleaves a luciferin substrate, specifically coelenterazine, emitting a photon to excite the fluorophore, causing emission of light. Since luciferase activity rate depends on the pH of the environment, the emission by the fluorophore is an indicator of pH, which can be calibrated or relatively measured by calculating the BRET ratio (475nm: 525nm recommended).
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This is the full plasmid construct of this part, viable in S. cerevisiae and E. coli.
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 16:55, 11 June 2018


bioluminescent "pHlash" reporter for intracellular pH

This is a genetically encodable probe that will report cytosolic pH in vivo of mammalian, yeast, or bacterial cells, through Bioluminescence Resonance Energy Transfer (BRET) data. The device utilizes a Rluc8 luciferase-cpVenus fluorophore fusion, where the Rluc8 luciferase protein exhibits pH-dependent activity. The luciferase cleaves a luciferin substrate, specifically coelenterazine, emitting a photon to excite the fluorophore, causing emission of light. Since luciferase activity rate depends on the pH of the environment, the emission by the fluorophore is an indicator of pH, which can be calibrated or relatively measured by calculating the BRET ratio (475nm: 525nm recommended).

This is the full plasmid construct of this part, viable in S. cerevisiae and E. coli.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 150
    Illegal AgeI site found at 759
    Illegal AgeI site found at 1648
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1500
    Illegal BsaI.rc site found at 1194
    Illegal SapI.rc site found at 1513