Difference between revisions of "Part:BBa K3225002"

 
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<partinfo>BBa_K3225002 short</partinfo>
 
<partinfo>BBa_K3225002 short</partinfo>
  
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This is a transcriptional factor that co-active promoter ExpR (BBa_K3225003) with AHL.
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Quorum sensing is a well-known cell-cell communication system to many bacteria as well as the plant pathogenic bacteria E.carotovora. The transcriptional factors ExpRs play the same role as LuxR in the E.carotovora quorum sensing systems. ExpRs are the receptors of OHHL (a quorum sensing signal of E. carotovora). After binding to OHHLs, ExpRs can positively or negatively regulate the protein expression. ExpR153(BBa K3225002), ExpR71(BBa K3225004), and EcbR(BBa K3225005) are the OHHL receptors in three E.carotovora subspecies. And all of the three receptors can bind a promoter pExp and regulate the downstream protein expression.
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Here we characterized the binding and induction features of three receptors of E. carotovora. The three receptors (ExpR153, ExpR71, and EcbR) were constitutive expressed under a constitutive promoter down streamed with the pExp regulating reporters (GFP).
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1)J23101- ExpR153 -pExp-GFP
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2)J23101- Exp71-pExp-GFP
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3)J23101- EcbR - pExp-GFP
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<partinfo>BBa_K3225002 parameters</partinfo>
 
<partinfo>BBa_K3225002 parameters</partinfo>
 
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==Result==
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we characterized the cellular sensing features within E.coli. The engineered bacteria were incubated in the plate reader. OD600 and fluorescent intensity were measured for cell density every 5 min for two hours. During this period, the inducer OHHL was added after 1 h.
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Before OHHL induction, it showed that there was no expression of the engineered bacteria with gene of ExpR71. However, for the bacteria with EcbR, it seemed that there was fluorescence output after 20 min incubation. And for ExpR153, the fluorescent protein was strongly expressed from the start (Fig. 1).
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<html><body><img style="width:800px;"src="https://static.igem.org/mediawiki/parts/3/3b/T--iBowu-China--Character-5003-fig1.png">
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Revision as of 19:16, 21 October 2019


ExpR153, transcriptional factor

Quorum sensing is a well-known cell-cell communication system to many bacteria as well as the plant pathogenic bacteria E.carotovora. The transcriptional factors ExpRs play the same role as LuxR in the E.carotovora quorum sensing systems. ExpRs are the receptors of OHHL (a quorum sensing signal of E. carotovora). After binding to OHHLs, ExpRs can positively or negatively regulate the protein expression. ExpR153(BBa K3225002), ExpR71(BBa K3225004), and EcbR(BBa K3225005) are the OHHL receptors in three E.carotovora subspecies. And all of the three receptors can bind a promoter pExp and regulate the downstream protein expression. Here we characterized the binding and induction features of three receptors of E. carotovora. The three receptors (ExpR153, ExpR71, and EcbR) were constitutive expressed under a constitutive promoter down streamed with the pExp regulating reporters (GFP).

1)J23101- ExpR153 -pExp-GFP

2)J23101- Exp71-pExp-GFP

3)J23101- EcbR - pExp-GFP


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 530
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 720
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 252


Result

we characterized the cellular sensing features within E.coli. The engineered bacteria were incubated in the plate reader. OD600 and fluorescent intensity were measured for cell density every 5 min for two hours. During this period, the inducer OHHL was added after 1 h. Before OHHL induction, it showed that there was no expression of the engineered bacteria with gene of ExpR71. However, for the bacteria with EcbR, it seemed that there was fluorescence output after 20 min incubation. And for ExpR153, the fluorescent protein was strongly expressed from the start (Fig. 1).