Regulatory

Part:BBa_K1328000

Designed by: Tianyi Li   Group: iGEM14_Tsinghua   (2014-10-09)
Revision as of 22:47, 17 October 2014 by Ted hou (Talk | contribs)

Part:BBa_K1328000 (HXT1-CMVmini) is a glucose-sensitive promoter of yeast origin and shown to function in both yeast and human cells. Exclusively activated by high glucose concentrations. 2014-HXT1%2BCMV-forget.png

Part HXT1+CMV is used as promoter for target gene under the regulation of glucose, consisting of HXT-1 promoter and CMV promoter.

HXT-1 is a low-affinity glucose transporter from Saccharomyces cerevisiae. There is a glucose response element in HXT-1 gene sensitive to glucose in human fibroblasts, which is used here as a glucose-dependent regulator.

Expression of target gene under the control of HXT1-CMV promoter is induced by high concentration of glucose.

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Figure 1. EGFP fluorescence observed in yeast cells cultured in a) %2 glucose, and b) 3% glycerol. c) Flow cytometry analysis: Cell count vs. EGFP intensity. Red: 2% glucose; Cyan: 3% glycerol.



Tsinghua_Project_Microbe_Fig_2.png Figure 2. Mean EGFP fluorescence measured by flow cytometry (FACS) in yeast cells, a) induced by different levels of glucose, and b) different incubation durations.


Tsinghua_Project_Microbe_Fig_3.png Figure 3. Flow cytometry analysis of glucose-induced EGFP expression in HEK293 cells cultured in various glucose concentrations and citrate (as non-glucose carbon source). Plotted is frequency vs. EGFP intensity.


For full characterization of this part, visit Team Tsinghua 2014's project wiki: http://2014.igem.org/Team:Tsinghua/Project/Microbe

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