Difference between revisions of "Part:BBa K3278002"
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However, expression leakage and delay are the two main problems of pdawn system. Leakage refers to the expression of target protein without light. According to our measurement, samples strictly kept away from light still had significant expression of GFP after being cultured for one night. Therefore, we carefully studied the gene circuits of pdawn system [2] [3] and hypothesized that enhancing the strength of the promoter of YF1 may enhance the repression of protein expression in the dark, thus solving the leakage problem.  
 
However, expression leakage and delay are the two main problems of pdawn system. Leakage refers to the expression of target protein without light. According to our measurement, samples strictly kept away from light still had significant expression of GFP after being cultured for one night. Therefore, we carefully studied the gene circuits of pdawn system [2] [3] and hypothesized that enhancing the strength of the promoter of YF1 may enhance the repression of protein expression in the dark, thus solving the leakage problem.  
  
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In this part, we use the modified tac promoter to substitute the lacIq promoter in front of YF1 and Fixk2, largely reducing the leakage while maintain reasonable expression under blue light.
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In this part, we use the modified tac promoter(abbreviated as mtac promoter) to substitute the lacIq promoter in front of YF1 and Fixk2, largely reducing the leakage while maintain reasonable expression under blue light.
  
 
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Revision as of 10:48, 19 October 2019


pDawn with modified tac promoter

pDawn is a blue light control system which can start the expression of target gene under the inducement of blue light. More information in BBa_K1075044 added by iGEM13_Bonn.

However, expression leakage and delay are the two main problems of pdawn system. Leakage refers to the expression of target protein without light. According to our measurement, samples strictly kept away from light still had significant expression of GFP after being cultured for one night. Therefore, we carefully studied the gene circuits of pdawn system [2] [3] and hypothesized that enhancing the strength of the promoter of YF1 may enhance the repression of protein expression in the dark, thus solving the leakage problem.

In this part, we use the modified tac promoter(abbreviated as mtac promoter) to substitute the lacIq promoter in front of YF1 and Fixk2, largely reducing the leakage while maintain reasonable expression under blue light.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2159
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 46
    Illegal NgoMIV site found at 178
    Illegal NgoMIV site found at 272
    Illegal NgoMIV site found at 565
    Illegal NgoMIV site found at 1059
    Illegal NgoMIV site found at 1077
    Illegal NgoMIV site found at 1167
    Illegal AgeI site found at 397
    Illegal AgeI site found at 1525
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1620
    Illegal BsaI.rc site found at 508