Difference between revisions of "Part:BBa K4156102"
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pLldR-Bxb1 consists of a fusion of the lactate-sensitive promoter pLldR and the serine integrase Bxb1. It will act as a complex regulatory for controlling downstream logic gates and transcription of genes.
 
pLldR-Bxb1 consists of a fusion of the lactate-sensitive promoter pLldR and the serine integrase Bxb1. It will act as a complex regulatory for controlling downstream logic gates and transcription of genes.
  
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===Characterization===
  
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==In vitro characterization and data analysis of the reported strains==
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To improve signaling stability as well as accuracy, we added Amplifying genetic switches based on serine integrase (Bxb1,TP901) to the R reporter(BBa_ ) to construct the AR reporter. Fig 1 indicates lactate (plldR) induced AR reporter with homogenized fluorescence intensity (mRFP/Cell). Comparing Fig1, 2, it can be seen that the fluorescence intensity of the AR reporter decreased significantly at a lactate concentration of 0 mM, and its expression was more stable over time. The fluorescence intensity of the AR reporter was also greater at other concentrations of lactate induction, and the difference between the fluorescence intensity after lactate induction at each concentration was more pronounced. This result indicates that the addition of amplifying genetic switch enhances the reporter intensity and robustness of the lactate biosensor.
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<figure style="text-align:center;">
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                <img style="max-width:700px;" src="https://static.igem.wiki/teams/4156/wiki/part/6-1.png" alt="control">
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                <figcaption><b>Figure 1:</b> Induction of downstream gene mRFP expression over time by the AR reporter consisting of plldR+Switch +mRFP at different lactate concentrations.</figcaption>
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              </figure>
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<html>
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<figure style="text-align:center;">
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                <img style="max-width:700px;" src="https://static.igem.wiki/teams/4156/wiki/part/3-1-6-2.png" alt="control">
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                <figcaption><b>Figure2:</b>Induction of downstream gene mRFP expression over time by the AR reporter consisting of plldR+mRFP at different lactate concentrations.</figcaption>
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              </figure>
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Revision as of 14:28, 10 October 2022


pLldR-Bxb1

pLldR-Bxb1 is constructed with L-lactate-sensitive promoter pLldR and serine integrase Bxb1, and is uesd for lactate-specific response for logic gate coupling.


Usage and Biology

pLldR-Bxb1 consists of a fusion of the lactate-sensitive promoter pLldR and the serine integrase Bxb1. It will act as a complex regulatory for controlling downstream logic gates and transcription of genes.

Characterization

In vitro characterization and data analysis of the reported strains

To improve signaling stability as well as accuracy, we added Amplifying genetic switches based on serine integrase (Bxb1,TP901) to the R reporter(BBa_ ) to construct the AR reporter. Fig 1 indicates lactate (plldR) induced AR reporter with homogenized fluorescence intensity (mRFP/Cell). Comparing Fig1, 2, it can be seen that the fluorescence intensity of the AR reporter decreased significantly at a lactate concentration of 0 mM, and its expression was more stable over time. The fluorescence intensity of the AR reporter was also greater at other concentrations of lactate induction, and the difference between the fluorescence intensity after lactate induction at each concentration was more pronounced. This result indicates that the addition of amplifying genetic switch enhances the reporter intensity and robustness of the lactate biosensor.

control
Figure 1: Induction of downstream gene mRFP expression over time by the AR reporter consisting of plldR+Switch +mRFP at different lactate concentrations.

control
Figure2:Induction of downstream gene mRFP expression over time by the AR reporter consisting of plldR+mRFP at different lactate concentrations.

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
    Illegal NheI site found at 1479
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1753
    Illegal XhoI site found at 1840
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 614
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2587