Difference between revisions of "Part:BBa K3350862"
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The key yqjF promoter was originally involved in the metabolism of aromatic compounds in bacteria and was later found to detect aromatic compounds in THT, DNT and other explosives. yqjF promoter detects the promoter's responsiveness to DNT and limit thresholds.
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Landmines pose a great threat to human lives and health. In our project, we designed a Bio-optical Landmine Detection device to achieve landmine detection with high sensitivity.
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We performed semirational mutation of the yqjF1st (BBa_K3350859) on the yqjF original sequence -35 region. Error-prone PCR was performed on the yqjF original sequence to obtain the yqjF2nd (BBa_K3350860). The mutation sites of the yqjF1st and yqjF2nd were accumulated to obtain the favorable mutants of the yqjF3rd generation.
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Our yqjF promoter (BBa_K1316002) was originally involved in the metabolism of aromatic compounds in bacteria and was later found to respond to chemicals, such as 2,4-dinitrotoluene (DNT) constantly released from landmines. However, previously reported detection limit of the yqjF promoter is as high as 25 mg/L of DNT; thus, we must improve the sensitivity of the yqjF promoter for its practical application.
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<img src="https://static.igem.org/mediawiki/parts/8/81/T--NEFU_China--ConversionPathway.png
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<img src="https://parts.igem.org/File:T--NEFU_China--yqjF%E9%98%88%E5%80%BC.png" style="width:50%;height=50%; ">
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<p>We optimized the response of the yqjF promoter to DNT in the following three aspects. We used semi-rational mutagenesis and error-prone PCR mutagenesis to randomly mutate the yqjF promoter and obtained its mutant versions yqjF1st and yqjF2nd (BBa_K3350859 and BBa_K3350860) with reduced DNT detection thresholds. After combining the favorable mutation sites of the yqjF1st and yqjF2nd, we generated the yqjF3rd promoter that show much reduced DNT detection threshold of 5 mg/L versus 25 mg/L of the wild-type yqjF promoter.
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Revision as of 12:08, 24 October 2020


yqjF3rd (promoter)
Landmines pose a great threat to human lives and health. In our project, we designed a Bio-optical Landmine Detection device to achieve landmine detection with high sensitivity.
Our yqjF promoter (BBa_K1316002) was originally involved in the metabolism of aromatic compounds in bacteria and was later found to respond to chemicals, such as 2,4-dinitrotoluene (DNT) constantly released from landmines. However, previously reported detection limit of the yqjF promoter is as high as 25 mg/L of DNT; thus, we must improve the sensitivity of the yqjF promoter for its practical application.

We optimized the response of the yqjF promoter to DNT in the following three aspects. We used semi-rational mutagenesis and error-prone PCR mutagenesis to randomly mutate the yqjF promoter and obtained its mutant versions yqjF1st and yqjF2nd (BBa_K3350859 and BBa_K3350860) with reduced DNT detection thresholds. After combining the favorable mutation sites of the yqjF1st and yqjF2nd, we generated the yqjF3rd promoter that show much reduced DNT detection threshold of 5 mg/L versus 25 mg/L of the wild-type yqjF promoter.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]