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Part:BBa_K4516019
hFoxO1-F.luciferase-R.luciferase:a platform to screen a small molecule FoxO1 antagonist
Profile
Name: hFoxO1-F.luciferase-R.luciferase
Base Pairs: 6187 bp
Origin: HepG2 cell genome
Properties: expression plasmid that can duplicate both in E.coli and HepG2 cells
Contribution
FoxO1, as a transcription factor, plays an important role in the regulation of blood glucose balance. FoxO1 in the liver can promote the expression of key gluconeogenesis enzyme genes, and an important mechanism of insulin regulation of blood sugar is to increase the phosphorylation of FoxO1, promote its nuclear export, and then reduce its transcriptional activation activity. According to this theory, we constructed a human FoxO1 full-length plasmid, used Firefly luciferase and Renilla luciferase as a reporter gene, established a FoxO1 transcriptional activation screening platform, and used this platform to screen out active compounds that inhibit FoxO1 transcriptional activation activity. In order to construct a FoxO1 expression plasmid that can shuttle both in E.coli and HepG2 cells, we designed the DNA sequences of hFoxO1 to be inserted into the XhoI and KpnI sites of the pcDNA3.1 vector (Fig.1), and transfected the HepG2 cells with the recombinant plasmid to set up our experiment platform.
Engineering Success
How we design our plasmid In order to construct a FoxO1 expression plasmid that can duplicate both in E.coli and HepG2 cells, we designed the DNA sequences of hFoxO1 to be inserted into the XhoI and KpnI sites of the pcDNA3.1 vector (Fig.1), and transfect the HepG2 cells with the recombinant plasmid and set up our experiment platform.
hFoxO1-F.luciferase-R.luciferase:a platform to screen a small molecule FoxO1 antagonist
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Unknown
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 783
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 4363
Illegal SapI.rc site found at 3273
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