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<p style="text-align:center"><b>Fig 3</b> Horizontal axis stands for different aromatic compounds utilized to induce HbpR biosensor. Vertical axis represents ON/OFF induction ratio of HbpR biosensor calculated from fluorescence intensity when exposed to corresponding aromatics. It is tested following <a href="http://2013.igem.org/Team:Peking/Team/Notebook/Protocols">Test Protocol 1</a>.   
 
<p style="text-align:center"><b>Fig 3</b> Horizontal axis stands for different aromatic compounds utilized to induce HbpR biosensor. Vertical axis represents ON/OFF induction ratio of HbpR biosensor calculated from fluorescence intensity when exposed to corresponding aromatics. It is tested following <a href="http://2013.igem.org/Team:Peking/Team/Notebook/Protocols">Test Protocol 1</a>.   
 
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Revision as of 06:22, 17 October 2013

J23114-HbpR-Terminator

For detailed information concerning HbpR, please visit 2013 Peking iGEM Biosensor HbpR


Characterization

In order to fine-tune the performance of HbpR biosensor, we constructed a library of Pc constitutive promoters at different intensity. BBa_K1031311 is composed of three elements, the constitutive Pc promoter J23114, coding sequence of HbpR and terminator B0015. (Fig 1)

Fig 1 Construction of biosensor circuit Pm/J23114-HbpR. Orange arrowheads represent promoters. RBSs are shown as green ovals and squares in dark red stand for B0015 terminator.


Sequence and features

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
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1735
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 449



Data shown

Construction and tuning

Induction ratio of HbpR biosensor library from ON/OFF test that utilizes different Pc promoters to control the expression of XylS. The dashed box refers to data for Pm/J23114-XylS biosensor circuit (Fig 2) This figure shows the biosensor circuit adopting J23114 obtains the optimal performance.

Fig 2 Horizontal axis stands for different HbpR biosensor with Pc promoters of different strength. The expression strength of these constitutive promoters, J23113, J23109, J23117, J23114 is 21, 106, 623 and 256, respectively, according to the Parts' registry. Two kinds of aromatics, namely 2-HBP and 2-ABP, shown with different color intensities, were tested following Test Protocol 1. Vertical axis represents the ON/OFF induction ratio. The dashed box refers to performance for Pm/J23114-XylS biosensor circuit.


Induction ratio of HbpR ON/OFF test when exposed to 78 aromatic compounds utilizing J23114 as constitutive promoter. It is shown that HbpR biosensor responds specific to 2-HBP and 2-ABP.

Fig 3 Horizontal axis stands for different aromatic compounds utilized to induce HbpR biosensor. Vertical axis represents ON/OFF induction ratio of HbpR biosensor calculated from fluorescence intensity when exposed to corresponding aromatics. It is tested following Test Protocol 1.


Dose-response curve of HbpR biosensor adopting J23114 when exposed to 2-HBP and 2-ABP.

Fig 4 Two inducers are shown in different color intensity. Horizontal axis stands for each inducer at different concentration. Vertical axis represents induction ratio calculated from fluorescence intensity, tested following Test Protocol 1.