Difference between revisions of "Part:BBa K2936009"

Revision as of 11:11, 18 October 2019

light-controlled repressible system

This part contains a light-controlled system and a repression system to achieve downstream gene expression under light conditions rather than working in dark conditions. In this system, under dark conditions, phosphorylated FixJ protein was transferred from YF1 protein to FixJ protein, and phosphorylated FixJ protein activated pFIXK2 promoter, then activated lacI protein expression. When the LacI protein binds to LacO, it prevents the downward expression of Lac promoter, the target gene is not activated. When cultured under light conditions, phosphorylation of FixJ protein was blocked and gene expression regulated by pfixk2 promoter was inhibited. Lac promoter expression was not inhibited, target gene expression was normal. If you want to use this system,you just need to link the target gene to the lactose operon gene and then introducing plasmid into the host cell.After that,If the transformant is successfully selected, it is possible to express the target gene by illumination.

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

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 <!-- Add more about the biology of this part here
 ===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.
 <!-- -->
 <span class='h3bb'>Sequence and Features</span>