Difference between revisions of "Part:BBa K1024002"
 
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<b>Design:</b> We constructed and improved standard BioBrick Parts about quorum sensing systems and modified the systems in prokaryotic microorganisms for usage in S. cerevisiae (Yeast). The LuxR gene is constitutively expressed, while the activation of Lux Promoter requires the signaling of N-Acyl Homoserine Lactone (AHL). Therefore, mCherry is activated in the presence of AHL.
 
<b>Design:</b> We constructed and improved standard BioBrick Parts about quorum sensing systems and modified the systems in prokaryotic microorganisms for usage in S. cerevisiae (Yeast). The LuxR gene is constitutively expressed, while the activation of Lux Promoter requires the signaling of N-Acyl Homoserine Lactone (AHL). Therefore, mCherry is activated in the presence of AHL.
 
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https://static.igem.org/mediawiki/igem.org/4/45/Tsinghua-BB-002.PNG
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<b>Figure 1. The construct of reporter for quorum sensing systems</b>
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<b>Test:</b> Yeasts after 8 hours of enlarge cultivation were divided into two three groups, induced by DMSO (control), 0.5uM AHL and 200uM AHL separately. After 0.5, 16 and 24 hours, yeasts were collected and tested for mCherry fluorescence by flow cytometry. Total fluorescence were obtained for each group. As shown in the figure, inducing with high concentration of 200uM AHL caused significant increase of mCherry fluorescence. Low concentration of AHL also induced higher fluorescence. Among the three tested time points, the fold change of fluorescence reached the highest when 16 hours after induction.
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https://static.igem.org/mediawiki/2013/d/d0/Tsinghua_Flow_Data_Final.jpg"
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<p><b>Figure 2. Fold change of mCherry fluorescence induced by AHL</b></p>
 
===Usage and Biology===
 
===Usage and Biology===
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Latest revision as of 20:51, 27 September 2013

Reporter for quorum sensing systems in yeast

Part Name: BBa_K1024002

Short Description: Reporter for quorum sensing systems in yeast

Part Type: Signaling

Design: We constructed and improved standard BioBrick Parts about quorum sensing systems and modified the systems in prokaryotic microorganisms for usage in S. cerevisiae (Yeast). The LuxR gene is constitutively expressed, while the activation of Lux Promoter requires the signaling of N-Acyl Homoserine Lactone (AHL). Therefore, mCherry is activated in the presence of AHL.

Tsinghua-BB-002.PNG

Figure 1. The construct of reporter for quorum sensing systems

Test: Yeasts after 8 hours of enlarge cultivation were divided into two three groups, induced by DMSO (control), 0.5uM AHL and 200uM AHL separately. After 0.5, 16 and 24 hours, yeasts were collected and tested for mCherry fluorescence by flow cytometry. Total fluorescence were obtained for each group. As shown in the figure, inducing with high concentration of 200uM AHL caused significant increase of mCherry fluorescence. Low concentration of AHL also induced higher fluorescence. Among the three tested time points, the fold change of fluorescence reached the highest when 16 hours after induction.

Tsinghua_Flow_Data_Final.jpg"

Figure 2. Fold change of mCherry fluorescence induced by AHL

Usage and Biology

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 1580
    Illegal BamHI site found at 2546
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 506
    Illegal BsaI site found at 893
    Illegal BsaI site found at 1280
    Illegal BsaI.rc site found at 547
    Illegal BsaI.rc site found at 934
    Illegal BsaI.rc site found at 1321