Difference between revisions of "Part:BBa M50439"

Latest revision as of 16:51, 11 June 2018

coding sequence 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 part is just the coding sequence for the fusion of the proteins, which are linked by the tri-amino acid sequence Ala-Glu-Leu.

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 204
Illegal AgeI site found at 1093
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 945
Illegal BsaI.rc site found at 639
Illegal SapI.rc site found at 958

@@ Line 3: / Line 3: @@
 <partinfo>BBa_M50439 short</partinfo>
-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.
+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 part is just the coding sequence for the fusion of the proteins, which are linked by the tri-amino acid sequence Ala-Glu-Leu.
 <!-- Add more about the biology of this part here