Difference between revisions of "Part:BBa K4779000"

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===Characterization===
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We treated the engineered yeast strain BY4741-pRS415-CMPG samples with a copper ion gradient, and detected them using a flow cytometer. The quantitative measurements of fluorescence intensity at various copper ion concentrations are shown in the graph. Within 200 uM, the sensor's signal output strength increases with an increase in copper ion concentration.
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Latest revision as of 18:38, 11 October 2023


CMPG:A yeast copper-induced reporter pathway with prm1 promoter

This is a copper ion-responsive signaling pathway in Saccharomyces cerevisiae based on MAPK system. When the cells were induced by copper ions, the circuit with prm1 promoter activates GFP expression to indicate the copper ion concentration.


Construction

We obtained the gene sequences of the basic parts from IGEM, such as the promoter of Cup1 (BBa_K945002), MFα2 (BBa_K110005), the promoter of prm1 (BBa_K1346004), green fluorescent protein (GFP) (BBa_K3112009), and the terminator CYC1 (BBa_K4278703), and sent them to GENEWIZ (Suzhou Genewiz Biotechnology Co., Ltd.) for synthesis. The new composite part (CMPG) (BBa_K4779000) is also synthesized.


nanjing-bioxstem-cmpg.png

Characterization

We treated the engineered yeast strain BY4741-pRS415-CMPG samples with a copper ion gradient, and detected them using a flow cytometer. The quantitative measurements of fluorescence intensity at various copper ion concentrations are shown in the graph. Within 200 uM, the sensor's signal output strength increases with an increase in copper ion concentration.


nanjing-bioxstem-cmpg2.png


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 279
    Illegal XbaI site found at 517
    Illegal SpeI site found at 343
    Illegal SpeI site found at 523
    Illegal PstI site found at 49
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 343
    Illegal SpeI site found at 523
    Illegal PstI site found at 49
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 55
    Illegal BamHI site found at 510
    Illegal XhoI site found at 8
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 279
    Illegal XbaI site found at 517
    Illegal SpeI site found at 343
    Illegal SpeI site found at 523
    Illegal PstI site found at 49
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 279
    Illegal XbaI site found at 517
    Illegal SpeI site found at 343
    Illegal SpeI site found at 523
    Illegal PstI site found at 49
    Illegal NgoMIV site found at 36
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2213
    Illegal SapI site found at 529