Revision as of 08:22, 19 October 2019

5*UAS-mRuby-hGluc-IL10

Usage and Biology

We construct a plasmid that can be controlled by blue light, it was composite with the 5xUAS-mRuby- P2A-hGluc. 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 10(IL-10), we wanted to use hGluc and mRuby to indicate the expression and secretion process of IL-10.

Figure1.the plasmid contain 5xUAS-mRuby-hGluc-IL10

Properties

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-10 expression with the control group. 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(with IL10 expression) number with light exposure

Figure3.cell number with light exposure of negative control group and experiment group

Sequence and Features