Regulatory

Part:BBa_K3350862

Designed by: Hao Xu   Group: iGEM20_NEFU_China   (2020-10-20)
Revision as of 12:08, 24 October 2020 by Hao-Xu (Talk | contribs)


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]


[edit]
Categories
//chassis/prokaryote/ecoli
//dna
Parameters
biology