Part:BBa_K3734001

CHREBP promoter

CHREBP is a promoter that can be induced by glucose.

1.Pattern diagram

Fig.1 The model diagram of CHREBP-LUC

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.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE 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
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE 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