Difference between revisions of "Part:BBa K3734026"

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.

Fig.1 The model diagram of LOV-VP16

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

Fig.2 Light controlled system testing experiment

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

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.

[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.