Difference between revisions of "Part:BBa K3734004"
 
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                 VP16
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                 GI
 
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                 <p>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
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                 <p>GI is a photosensitive protein, under the exposure of 450nm blue light, its structure will alter and can form a dimer with LOV. We used GI as a part of our blue light switch.
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                    combine with UAS, to locate UAS.
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                 <img class="image" src="https://2021.igem.org/wiki/images/f/f3/T--CSU_CHINA--tupian8.png">
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                    <img width="400px" src="https://2021.igem.org/wiki/images/8/87/T--CSU_CHINA--tupian5.png"></p>
 
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                 <p style="width: 80%;text-align:center;font-size: .9rem; margin: -1rem auto 1rem auto; color: #888;">Fig.1 The model diagram of LOV-VP16</p>
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                 <p style="width: 80%;text-align:center;font-size: .9rem; margin: -1rem auto 1rem auto; color: #888;">Fig.1 The model diagram of GIP-miR21T-GI-GAL4-4XmiR21TC</p>
 
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                 <p>GAL4 combining domain is universal among eukaryotic cells, but the activation factor needs to be chosen differently according to the chassis creature.
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                 <p>The duration of the combination of GI and LOV after the blue light exposure will alter as the illuminate method and the intensity of the blue light differ. We used strong light for a 30min exposure and then use weak light for another 30min
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                    exposure.
 
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                 <p>[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.
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                 <p>[1]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.
 
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Latest revision as of 08:16, 20 October 2021

GI

GI 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 LOV.

GI

GI is a photosensitive protein, under the exposure of 450nm blue light, its structure will alter and can form a dimer with LOV. We used GI as a part of our blue light switch.

1.Pattern diagram

Fig.1 The model diagram of GIP-miR21T-GI-GAL4-4XmiR21TC

2.Caution

The duration of the combination of GI and LOV after the blue light exposure will alter as the illuminate method and the intensity of the blue light differ. We used strong light for a 30min exposure and then use weak light for another 30min exposure.

Reference：

[1]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
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 3147
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 3042
    Illegal XhoI site found at 695
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1197
    Illegal AgeI site found at 2424
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1821