Difference between revisions of "Part:BBa K4115040"
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This composite part is used to produce 3OC6 HSL by LacI [https://parts.igem.org/Part:BBa_C0161 (BBa_C0161)] under a constitutive promoter J23101[https://parts.igem.org/Part:BBa_J23101 (BBa_J23101)] and send it to the receiver protein LuxR [https://parts.igem.org/Part:BBa_C0062 (BBa_C0062)]in the other <i>E. coli</i>.When the LuxR receives the signal, it will regulate and express the downstream report gene sfGFP, thus we can detect whether LuxI is functional by observing the fluorescence intensity under UV light.
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This composite part is used to produce 3OC6 HSL by LacI [https://parts.igem.org/Part:BBa_C0161 (BBa_C0161)][1] under a constitutive promoter J23101[https://parts.igem.org/Part:BBa_J23101 (BBa_J23101)] and send it to the receiver protein LuxR [https://parts.igem.org/Part:BBa_C0062 (BBa_C0062)]in the other <i>E. coli</i>.When the LuxR receives the signal, it will regulate and express the downstream report gene sfGFP [2], thus we can detect whether LuxI is functional by observing the fluorescence intensity under UV light.
  
 
==Proof of function==
 
==Proof of function==
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===Usage and Biology===
 
  
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K4115040 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4115040 SequenceAndFeatures</partinfo>
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== References ==
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<small>
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[1] Daer R, Barrett CM, Melendez EL, Wu J, Tekel SJ, Xu J, Dennison B, Muller R, Haynes KA. Characterization of diverse homoserine lactone synthases in Escherichia coli. PLoS One. 2018 Aug 23;13(8):e0202294. doi: 10.1371/journal.pone.0202294. PMID: 30138364; PMCID: PMC6107141.
  
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[2] Kylilis, N., Tuza, Z.A., Stan, GB. et al. Tools for engineering coordinated system behaviour in synthetic microbial consortia. Nat Commun 9, 2677 (2018). https://doi.org/10.1038/s41467-018-05046-2
  
 
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Revision as of 07:15, 4 October 2022


J23101-B0034-LuxI-B0015

J23101-B0034-LuxI-B0015
Function gene regulator
Use in E. coli
RFC standard RFC 10, RFC 10 compatible
Backbone pBBR1
Submitted by ShanghaiTech_China

This composite part is used to produce 3OC6 HSL by LacI (BBa_C0161)[1] under a constitutive promoter J23101(BBa_J23101) and send it to the receiver protein LuxR (BBa_C0062)in the other E. coli.When the LuxR receives the signal, it will regulate and express the downstream report gene sfGFP [2], thus we can detect whether LuxI is functional by observing the fluorescence intensity under UV light.

Proof of function

We encapsulated E. coli with this composite part and E. coli with LuxR and sfGFP into alginate gel beads. The E. coli with LuxI secreted the 3OC6 HSL signal molecule, and the E. coli with LuxR received the signal and regulate the expression of downstream sfGFP, making it express. So we can detect green fluorescence in the latter E. coli under the irradiation of ultraviolet light.

Figure 1: sfGFP is expressed under the regulation of LuxR between E. coli in a single gel bead. Two parallel groups are given and the above two are control groups.
























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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

References

[1] Daer R, Barrett CM, Melendez EL, Wu J, Tekel SJ, Xu J, Dennison B, Muller R, Haynes KA. Characterization of diverse homoserine lactone synthases in Escherichia coli. PLoS One. 2018 Aug 23;13(8):e0202294. doi: 10.1371/journal.pone.0202294. PMID: 30138364; PMCID: PMC6107141.

[2] Kylilis, N., Tuza, Z.A., Stan, GB. et al. Tools for engineering coordinated system behaviour in synthetic microbial consortia. Nat Commun 9, 2677 (2018). https://doi.org/10.1038/s41467-018-05046-2