Part:BBa_K3734012

TRE

TRE is a DNA structure domain that combined with TetR.

TRE

TRE is a DNA structure domain that combined with TetR. It is a part of Tet-off system. Our first purpose of using it is to make phosphated ELK1 to locate and activate the expression of downstream genes (miRNA). The second reason we used it is because of the Tet-off system consists of TetR and TRE can be blocked by tetracycline. Tetracycline will stop the combination between TetR and TRE, stop the system from working. This provides a mandatory brake system for our loop. We can make sure the security and prevent the hypoglycemia caused by abnormal feedback through exogenous tetracycline.

1.Pattern diagram

Fig.1 The model diagram of TRE's work

2.1 Method

Phosphated ELK will activate the expression of downstream report gene mCherry after the combination of TRE and TetR. We used lase confocal microscope to observe the expression.

2.2 Results

TRE works well.

Fig.2 Under Laser confocal microscopy, fluorescence of mCherry expression downstream of Tet-Off system

3.Caution

If the green fluorescent protein is expressed too much, and if the green fluorescence is too strong, it may cause a series of colors. In order to ensure the accuracy of the experimental results, the level of noise reduction should be raised.

Reference：

[1]Haifeng Ye, Mingqi Xie, Shuai Xue.Self-adjusting synthetic gene circuit for correcting insulin resistance[J].Nat Biomed Eng.2017 Jan;1(1):0005.

Sequence and Features

Assembly Compatibility:

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