Difference between revisions of "Part:BBa K2986009"
 
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We construct a plasmid that can be controlled by blue light, it was composite with the 5xUAS-mRuby-hGluc-IL8. We used a light-switch transactivator named GVAPO, which can bind to promoter and initiates transcription of upstream gene in a short time.
 
We construct a plasmid that can be controlled by blue light, it was composite with the 5xUAS-mRuby-hGluc-IL8. 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 (cytokine secretion), and to obtain better indicators for long-term observation, we need to replace cytokines with Humanized Gaussia luciferase (hGluc). P2A is 2A peptide allows an open reading frame (ORF) to translate a peptide chain into several independent peptide chains. we added the target gene Interleukin 8(IL-8), we wanted to use hGluc and mRuby to indicate the expression and secretion process of IL-8.
 
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 (cytokine secretion), and to obtain better indicators for long-term observation, we need to replace cytokines with Humanized Gaussia luciferase (hGluc). P2A is 2A peptide allows an open reading frame (ORF) to translate a peptide chain into several independent peptide chains. we added the target gene Interleukin 8(IL-8), we wanted to use hGluc and mRuby to indicate the expression and secretion process of IL-8.
−
[[File:T-SUSTECH ZHILI.png|450px|thumb|center|Figure1.plasmid with 5xUAS-mRuby-hGluc-IL8]]
 
  
  
 
<h2>Properties</h2>
 
<h2>Properties</h2>
 +
 +
[[File:T-SUSTECH ZHILI.png|450px|thumb|center|Figure1.plasmid with 5xUAS-mRuby-hGluc-IL8]]
 +
 
We used hGluc as a long-term indicator for the target gene expression, and we get the data after 48 hrs blue light exposure. We contrasted IL-8 expression with the control group.
 
We used hGluc as a long-term indicator for the target gene expression, and we get the data after 48 hrs blue light exposure. We contrasted IL-8 expression with the control group.
 
we can see from the data that IL-8 transcription process is influenced by the light exposure time.
 
we can see from the data that IL-8 transcription process is influenced by the light exposure time.

Latest revision as of 08:37, 20 October 2019


5*UAS-mRuby-hGluc-IL8

Usage and Biology

We construct a plasmid that can be controlled by blue light, it was composite with the 5xUAS-mRuby-hGluc-IL8. 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 (cytokine secretion), and to obtain better indicators for long-term observation, we need to replace cytokines with Humanized Gaussia luciferase (hGluc). P2A is 2A peptide allows an open reading frame (ORF) to translate a peptide chain into several independent peptide chains. we added the target gene Interleukin 8(IL-8), we wanted to use hGluc and mRuby to indicate the expression and secretion process of IL-8.


Properties

Figure1.plasmid with 5xUAS-mRuby-hGluc-IL8

We used hGluc as a long-term indicator for the target gene expression, and we get the data after 48 hrs blue light exposure. We contrasted IL-8 expression with the control group. we can see from the data that IL-8 transcription process is influenced by the light exposure time.

Figure2.IL-8 mRNA expression amount under blue light exposure

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 BamHI site found at 906
    Illegal XhoI site found at 49
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 189
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1527
    Illegal SapI.rc site found at 210