Difference between revisions of "Part:BBa K4516001"
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How we design our plasmid | 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. | 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. | ||
| − | [[File:T-- | + | [[File:T--Jiangsu United--BBa K4516019-figure1.png|500px|thumb|center|Fig. 1 The map of recombinant plasmid pcDNA3.1-hFoxO1..]] |
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Revision as of 08:59, 26 September 2022
hFoxO1
hFoxO1
Profile
Name: hFoxO1 Origin: HepG2 cell genome
Contribution
Forkhead box protein (Fox) is a family of transcription factors. The DNA binding region of this family of proteins has a conserved winglike helix structure. There are currently 17 subfamilies in this family, of which the FoxO subfamily is the most well studied. There are four subtypes: FoxO1, FoxO3, FoxO4, and FoxO6 in mammals. FoxO1 has four domains, which are DNA-binding domain, nuclear localization domain, nuclear export sequence, and transcriptional activation domain. It binds with IRE sequence and plays a role in regulating downstream genes. FoxO1 is mainly expressed in insulin-responsive tissues, The main role of FoxO1 is to regulate downstream target genes, such as PEPCK, G6Pase, PGC1-α, and PDK-4 to promote gluconeogenesis and can regulate cell proliferation, gluconeogenesis, and energy metabolism.
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.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 289
Illegal NotI site found at 298 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 554
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]