Difference between revisions of "Part:BBa K909008"

 
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<partinfo>BBa_K909008 short</partinfo>
 
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Tetracycline repressor DNA binding domain (<partinfo>BBa_K909007</partinfo>) fusion with the N-terminus of the truncated version of UVR8 (<partinfo>BBa_K909009</partinfo>).
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Tetracycline repressor DNA binding domain fusion with truncated version of UVR8 (tetR-DBD-UVR8).
 
Tetracycline repressor DNA binding domain fusion with truncated version of UVR8 (tetR-DBD-UVR8).
 
UVR8 is a UV-B sensing receptor in plants necessary for UV-B protection. In dark state UVR8 forms a dimer which is monomerized by the UV-B light (280-315 nm). A truncated version of UVR8 (14-440 amino acids) was fused with tetR DNA binding domain (tetR-DBD), lacking of dimerization domain. Such mutant shows no repression of Ptet promoter, however fusion with dimerizing protein (UVR8) returns tetR-DBD activity. Later UV-B exposure breaks UVR8 dimer and releases tetR-DBD-UVR8 from Ptet, activating transcription. Thus, this tetR-DBD-UVR8 can be used as a one step UV-B on switch.  
 
UVR8 is a UV-B sensing receptor in plants necessary for UV-B protection. In dark state UVR8 forms a dimer which is monomerized by the UV-B light (280-315 nm). A truncated version of UVR8 (14-440 amino acids) was fused with tetR DNA binding domain (tetR-DBD), lacking of dimerization domain. Such mutant shows no repression of Ptet promoter, however fusion with dimerizing protein (UVR8) returns tetR-DBD activity. Later UV-B exposure breaks UVR8 dimer and releases tetR-DBD-UVR8 from Ptet, activating transcription. Thus, this tetR-DBD-UVR8 can be used as a one step UV-B on switch.  
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===Usage and Biology===
 
===Usage and Biology===
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TetR-DBD (1-127 amino acids of original TetR) unable to bind DNA and repress transcription from TetR responsive promoter Ptet due to the lacking dimerization domain. Nevertheless, fusions with dimerizing proteins restores TetR-DBD DNA binding activity.
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UVR8 is a plant UV-B photoreceptor, which switches from dimeric to monomeric form in the presence of UV-B.  By fusing UVR8 with DNA binding domain of TetR, we can regulate gene expression with UV-B light from Ptet (figure 1).
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[[Image:Uvr8uv.jpg|frameless|200px|center|thumb| Figure 1. UV-B light induced derepression by TetR-DBD-UVR8]
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Revision as of 02:28, 27 September 2012

tetR-DBD-UVR8 fusion protein

Tetracycline repressor DNA binding domain (BBa_K909007) fusion with the N-terminus of the truncated version of UVR8 (BBa_K909009).


Usage and Biology

TetR-DBD (1-127 amino acids of original TetR) unable to bind DNA and repress transcription from TetR responsive promoter Ptet due to the lacking dimerization domain. Nevertheless, fusions with dimerizing proteins restores TetR-DBD DNA binding activity.

UVR8 is a plant UV-B photoreceptor, which switches from dimeric to monomeric form in the presence of UV-B. By fusing UVR8 with DNA binding domain of TetR, we can regulate gene expression with UV-B light from Ptet (figure 1).


Figure 1. UV-B light induced derepression by TetR-DBD-UVR8]


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 902
    Illegal PstI site found at 1556
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 418
    Illegal PstI site found at 902
    Illegal PstI site found at 1556
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 382
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 902
    Illegal PstI site found at 1556
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 902
    Illegal PstI site found at 1556
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 433