Difference between revisions of "Part:BBa K2257007"
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FrmR can form DNA-protein cross-links with a special promotor PfrmAB to prevent activating downstream gene transcription.<br/>
 
FrmR can form DNA-protein cross-links with a special promotor PfrmAB to prevent activating downstream gene transcription.<br/>
 
Formaldehyde is able to covalently modify protein FrmR, inhibiting its function, so when formaldehyde exists, promotor PfrmAB is capable of activating downstream gene transcription by the destruction of the promoter PfrmAB- the protein FrmR cross-link.</p>
 
Formaldehyde is able to covalently modify protein FrmR, inhibiting its function, so when formaldehyde exists, promotor PfrmAB is capable of activating downstream gene transcription by the destruction of the promoter PfrmAB- the protein FrmR cross-link.</p>
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[[File:T-Nanjing-China-ch2o-l2.png|600px|thumb|center]Figure 1. Optical density(600nm) of (a) Escherichia coli BL21 and (b) recombinant bioluminescent Escherichia coli BL21 harboring frmR-RFP fusion after 10 hours’ incubation with 800uM different aldehydes]
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[[File:T-Nanjing-China-ch2o-l2.png|600px|thumb|center|Figure 1. Optical density(600nm) of (a) Escherichia coli BL21 and (b) recombinant bioluminescent Escherichia coli BL21 harboring frmR-RFP fusion after 10 hours’ incubation with 800uM different aldehydes]]
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[[File:T-Nanjing-China-ch2o-l1.png|500px|thumb|center]Figure 2. Response growth curve for recombinant bioluminescent Escherichia coli BL21 to different concentration of formaldehyde]
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[[File:T-Nanjing-China-ch2o-l1.png|500px|thumb|center|Figure 2. Response growth curve for recombinant bioluminescent Escherichia coli BL21 to different concentration of formaldehyde]]
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[[File:T-Nanjing-China-ch2o-l3.png|500px|thumb|center]Figure 3. Fluorescence test of various aldehydes using recombinant bioluminescent Escherichia coli BL21]
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[[File:T-Nanjing-China-ch2o-l3.png|500px|thumb|center|Figure 3. Fluorescence test of various aldehydes using recombinant bioluminescent Escherichia coli BL21]]
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[[File:T-Nanjing-China-ch2o-l4.png|500px|thumb|center]Figure 4. The tolerance of recombinant bioluminescent Escherichia coli BL21 to various concentration of formaldehyde]
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[[File:T-Nanjing-China-ch2o-l4.png|500px|thumb|center|Figure 4. The tolerance of recombinant bioluminescent Escherichia coli BL21 to various concentration of formaldehyde]]
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[[File:T-Nanjing-China-ch2o-l5.png|500px|thumb|center]Figure 5. Response fluorescence intensity for recombinant bioluminescent Escherichia coli BL21 to formaldehyde]
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[[File:T-Nanjing-China-ch2o-l5.png|500px|thumb|center|Figure 5. Response fluorescence intensity for recombinant bioluminescent Escherichia coli BL21 to formaldehyde]]
 
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Revision as of 14:26, 30 October 2017


This part is used for the detection of formaldehyde.

Formaldehyde sensor sequence


Usage and Biology

This protein FrmR acts as regulator of protein FrmAB. FrmA and FrmB are two proteins that are participating in the detoxication of formaldehyde in E.coli.
FrmR can form DNA-protein cross-links with a special promotor PfrmAB to prevent activating downstream gene transcription.
Formaldehyde is able to covalently modify protein FrmR, inhibiting its function, so when formaldehyde exists, promotor PfrmAB is capable of activating downstream gene transcription by the destruction of the promoter PfrmAB- the protein FrmR cross-link.

Figure 1. Optical density(600nm) of (a) Escherichia coli BL21 and (b) recombinant bioluminescent Escherichia coli BL21 harboring frmR-RFP fusion after 10 hours’ incubation with 800uM different aldehydes
Figure 2. Response growth curve for recombinant bioluminescent Escherichia coli BL21 to different concentration of formaldehyde
Figure 3. Fluorescence test of various aldehydes using recombinant bioluminescent Escherichia coli BL21
Figure 4. The tolerance of recombinant bioluminescent Escherichia coli BL21 to various concentration of formaldehyde
Figure 5. Response fluorescence intensity for recombinant bioluminescent Escherichia coli BL21 to formaldehyde

] Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 226
    Illegal XhoI site found at 462
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 520

The sequence is composed of PfrmR, gene frmR, flag tag, PfrmAB, gene RFP.
When the pathway works, we can see that the E.coli turns to red with naked eyes at the presence of formaldehyde.
The fluorescence intensity of protein RFP and the depth of the color in naked eyes are both in positive correlation with the concentration of formaldehyde within the tolerable range.