Part:BBa_K2986011

5*UAS-mRuby

Usage and Biology

We construct a plasmid that can be controlled by blue light, it was composite with the 5xUAS-mRuby. We used a light-switch transactivator named GVAPO, which can bind to promoter and initiates transcription of upstream gene in a short time. And 5xUASis a site for GVAPO binding and perform its function. After light activation, GVAP homodimerizes and interacts with this sequence to initiates expression of gene interest. We use mRuby to be a reporter of our target gene expression, but we were not sure whether mRuby itself can affect gene expression. So we constructed the 5xUAS-mRuby to be a negative control.

Figure1.plasmid with 5xUAS-mRuby

Properties

The data showed that inside Hela cell, the mRuby indicator system has little influence on cytokine expression and secretion. This showed that the Characterization of system was reliable.

Figure2.cell number with light exposure of negative control group and experiment group

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 120
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 167
  Illegal XhoI site found at 49
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 207