Reporter

Part:BBa_K4226000

Designed by: Siyu Han   Group: iGEM22_CAFA_China   (2022-09-14)
Revision as of 06:34, 26 September 2022 by Tigress K (Talk | contribs) (→‎Characterized by CAFA_China 2022)


3WJ-Bro

3WJ-Bro is used as a protein- independent reporter system based on a fluorescent RNA aptamer. It is applied as a gene marker in creating a system for reporting the presence and expression of target genes. The 3WJ-Bro can be used to ligate fluorescent aptamers to examine the relE RNA at transcriptional level in Escherichia coli cells, obviating the need for accumulation of foreign proteins.

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]


Characterized by CAFA_China 2022

  • We designed the genetic circuit that contains sulAp (sulA promoter), Amp30E, relE gene (RelE toxin), mScarlet-I (mSarlet-I) and 3WJ-Bro, and transferred it into DH10B competent cells (The control group did not contain the Amp30E Amplification Device).
  • After bacterial culture and UV induction, we measured the OD600 and fluorescence values under 485/510nm. Then, the fluorescence per OD was calculated.
  • With the lengthening of culture time, the RNA synthesis of relE increased gradually in the experimental group. According to the results, the 3WJ-Bro can be used to ligate fluorescent aptamers to examine the relE RNA synthesis at transcriptional level.
Figure: 3WJ-Bro was ligated with fluorescent aptamers in order to examine the level of relE RNA synthesis (485/510nm). Control group: pSB1C3-sulAp-relE-mSarlet-l-3WJxbro; experimental group: pSB1C3-sulAp-Amp30E-relE-mSarlet-l-3WJxbro.



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