Part:BBa_K3734026

LOV-VP16

LOV is a kind of light-activated protein. If you shine blue light on it, it will change its own structure and will be able to combined with GI.VP16 is a kind of protein that can activate downstream gene expression.When you shine blue light on the cell, LOV will combined with GI, and GAL4 will combined with 9XUAS.Then VP16 will activate downstream gene expression.

LOV-VP16

LOV is a photosensitive protein, the structure will alter under 450nm blue light exposure and form a dimer with GI. We used LOV as a part of our blue light switch. VP16 is a transcription activation factor, also commonly used in double yeast hybrid system. It can realize the activation of downstream UAS transcription. It is usually used together with GAL4 combining domain, which can recognize and combine with UAS, to locate UAS.

1.Pattern diagram

Fig.1 The model diagram of LOV-VP16

2.1 Method

We used report gene LUC to represent the effect of LOV-IP16

2.2 Result

Fig.2 Light controlled system testing experiment

Fig.3 Expression level analysis under blue light irradiation and dark treatment

3.Caution

The duration of GI and LOV after 450nm blue light exposure will vary with the irradiation method, intensity, etc. In the experiment, we adopted the method of irradiating strong light for 30 minutes first, and then low light for 30 minutes. The GAL4 binding domain is universal in eukaryotic cells, but transcription activators (such as VP16) that cooperate with GAL4 need to be properly selected according to different chassis organisms.

Reference：

[1]Chun Jeih Ryu , Charles E Whitehurst, Jianzhu Chen.Expression of Gal4-VP16 and Gal4-DNA binding domain under the control of the T lymphocyte-specific lck proximal promoter in transgenic mice[J].BMB Rep. 2008 Aug 31;41(8):575-80.

[2]Masayuki Yazawa , Amir M Sadaghiani, Brian Hsueh.Induction of protein-protein interactions in live cells using light[J].Nat Biotechnol. 2009 Oct;27(10):941-5.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 293
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]