Difference between revisions of "Part:BBa K2936009"
 
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===Usage and Biology===
 
===Usage and Biology===
 
This part contains a light-controlled system and a repression system to achieve the regulations of target gene expression under light conditions. If you want to use this system, you just need to insert the target gene after the Lac operator in the plasmid, which was further transferred into the host cell.
 
This part contains a light-controlled system and a repression system to achieve the regulations of target gene expression under light conditions. If you want to use this system, you just need to insert the target gene after the Lac operator in the plasmid, which was further transferred into the host cell.
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We link the repression system of the lactose operon directly behind the original light-controlled system (behind the GFP).In this system, under dark conditions, phosphate group of YF1 protein will be transferred to FixJ protein, and the phosphorylated FixJ protein will initiate the activation of FIXK2 promoter to drive the expression of downstream gene including eGFP and LacI. When the LacI protein was expressed, it will prevent the function of Lac promoter. As a result, the downstream target gene will not be expressed. When induced by light, phosphorylation of FixJ protein was blocked and gene expression regulated by pfixk2 promoter was inhibited. Lac promoter expression was not inhibited, downstream gene will be expressed.
 
We link the repression system of the lactose operon directly behind the original light-controlled system (behind the GFP).In this system, under dark conditions, phosphate group of YF1 protein will be transferred to FixJ protein, and the phosphorylated FixJ protein will initiate the activation of FIXK2 promoter to drive the expression of downstream gene including eGFP and LacI. When the LacI protein was expressed, it will prevent the function of Lac promoter. As a result, the downstream target gene will not be expressed. When induced by light, phosphorylation of FixJ protein was blocked and gene expression regulated by pfixk2 promoter was inhibited. Lac promoter expression was not inhibited, downstream gene will be expressed.
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Latest revision as of 11:28, 18 October 2019


light-controlled repressible system

Usage and Biology

This part contains a light-controlled system and a repression system to achieve the regulations of target gene expression under light conditions. If you want to use this system, you just need to insert the target gene after the Lac operator in the plasmid, which was further transferred into the host cell.

We link the repression system of the lactose operon directly behind the original light-controlled system (behind the GFP).In this system, under dark conditions, phosphate group of YF1 protein will be transferred to FixJ protein, and the phosphorylated FixJ protein will initiate the activation of FIXK2 promoter to drive the expression of downstream gene including eGFP and LacI. When the LacI protein was expressed, it will prevent the function of Lac promoter. As a result, the downstream target gene will not be expressed. When induced by light, phosphorylation of FixJ protein was blocked and gene expression regulated by pfixk2 promoter was inhibited. Lac promoter expression was not inhibited, downstream gene will be expressed.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 605
    Illegal NgoMIV site found at 677
    Illegal NgoMIV site found at 767
    Illegal NgoMIV site found at 785
    Illegal NgoMIV site found at 1297
    Illegal NgoMIV site found at 1590
    Illegal NgoMIV site found at 1684
    Illegal AgeI site found at 319
    Illegal AgeI site found at 1465
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1354
    Illegal BsaI.rc site found at 218