Difference between revisions of "Part:BBa K3734001"
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| + | ===Usage and Biology=== | ||
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| + | <span class='h3bb'>Sequence and Features</span> | ||
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| + | ===Functional Parameters=== | ||
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Revision as of 13:11, 19 October 2021
CHREBP promoter
CHREBP promoter is a glucose-induced promoter, and its expression increases with the increase of glucose concentration.
CHREBP promoter
1.Pattern diagram
<img class="image" src="">
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.
<img class="image" src="">
Fig.2 Electrophoretic diagram of CHREBP PCR product
<img class="image" src="">
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.
<img class="image" src="">
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.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 584
Illegal BamHI site found at 346
Illegal XhoI site found at 449
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
Illegal BsaI site found at 3632
Illegal BsaI.rc site found at 452
Illegal BsaI.rc site found at 475
Illegal BsaI.rc site found at 2160
Illegal BsaI.rc site found at 2270
Illegal SapI.rc site found at 3150