Difference between revisions of "Part:BBa K3734001"
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Latest revision as of 08:17, 21 October 2021
CHREBP promoter
CHREBP is a promoter that can be induced by glucose.
CHREBP promoter
1.Pattern diagram
Fig.1 The model diagram of CHREBP-LUC
2.Experiment
2.1 Method
We used LUC as report genes to reflect the level of expression through detecting luminescence value at 560nm wavelength and the error REN luminescence caused by the number of eliminated cells.
2.2 Result
CHERBP promoter is induced by glucose and the expression rises along with the raise of glucose concentration.
Fig.2 Electrophoretic diagram of CHREBP PCR product
Fig.3 The expression level of inducible promoter CHREBP was respectively analyzed at 48h in 25mM, 5.6mm and 0mM glucose culture
To eliminate the effect of residual glucose in the medium during transmission, we have conducted experiments in a 72-hour group, the result fits our anticipations more.
Fig.4 The expression level of inducible promoter CHREBP was respectively analyzed at 72h in 25mM, 5.6mm and 0mM glucose culture
3.Caution
Due to the relatively long sequence of CHREBP promoter, DH5α receptor cells containing recombinant enzymes are difficult to meet the
requirements during cloning, and receptor cells with recombinant enzymes may
need to be removed.
Meanwhile, we hope that we can shorten the length of the sequence to improve it although it can work quite well.
Reference:
[1]Jian Meng, Ming Feng, Weibing Dong.Identification of HNF-4α as a key transcription factor to promote ChREBP expression in response to glucose[J].Sci Rep. 2016 Mar 31;6:23944.
- 10COMPATIBLE WITH RFC[10]
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- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 584
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Illegal XhoI site found at 2640 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 3317
Illegal BsaI site found at 3385
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Illegal BsaI.rc site found at 452
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