Composite

Part:BBa_K4169021

Designed by: Rui Tian   Group: iGEM22_HZAU-China   (2022-09-30)
Revision as of 14:05, 12 October 2022 by Tery (Talk | contribs)


pFrmR+FrmRAB - An operon that degrades formaldehyde



Basic Description

PfrmR is found upstream of the frmRAB formaldehyde detoxification operon. FrmR, the first product of the operon, is a member of the DUF156 family of DNA-binding transcriptional regulators. It binds the frmRAB promoter region and is negatively allosterically modulated by formaldehyde. FrmR is specific to formaldehyde. The genes frmA and frmB encode a formaldehyde dehydrogenase and S-formylglutathione hydrolase, respectively, and are responsible for detoxifying formaldehyde to formic acid in a glutathione-dependent pathway. The negative-feedback regulation of the frmRAB operon is similar to that of many other prokaryotic operons, whereby the transcription factor represses its own transcription.

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Figure 1.Without formaldehyde


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Figure 2.Containing formaldehyde

Origin

E.coli K12 MG1655.

Biology characteristic

1.This operon is induced by formaldehyde and degrades formaldehyde.

2.The protein expressed by FrmR can inhibit the upstream promoter, but formaldehyde can bind to frmR to prevent the inhibitory effect.

Experimental verification

Experiment 1

Culture:LB liquid medium +40%HCHO saturated aqueous solution.

Instrument: microplate reader.

The experimental group:DH5α with pFrmR+FrmRAB.

The control group:DH5α(No plasmid transformation).




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 242
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 679
    Illegal AgeI site found at 1003
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None