Device

Part:BBa_K4226004

Designed by: Siyu Han   Group: iGEM22_CAFA_China   (2022-09-19)
Revision as of 05:42, 27 September 2022 by Tigress K (Talk | contribs)


Amp30E Amplification Device

The Amp30E Amplification Device is an amplifier module includes hrpR Gene, hrpS Gene and hrpL Promoter, this engineered multi-layered transcriptional amplifiers can be able to sequentially boost output expression level of target genes.


Usage and Biology

Characterized by CAFA_China 2022

  • We applied a typical cellular sensor that can be abstracted as a three-stage processor comprising a sensing module that recognizes and transduces external signals into intracellular transcriptional signals, a computing module that modulates the transduced sensor signals, and an output actuating module that executes physiological responses.
  • We first used the promoter of sulA as sensor module. The amplifier module (Amp30E Amplification Device) includes hrpR, hrpS and PhrpL , and the eGFP was used as an output module to test the effect of amplifier module. The gene circuits is shown bellow:
Gene circuit: PSB1C3-sulAp-Amp30E-eGFP

  • The recombinant cells were cultured and the parameters were measured using microplate reader. The fluorescence per OD significantly increased after induction with different times of UV exposure (UVC, 254nm) . The results are as follows:
Figure: The result of eGFP expression (Fluorescence per OD) after irradiated under UVC (254nm).

  • The results showed that the Amp30E Amplification Device (PSB1C3-sulAp-Amp30E-eGFP) significantly increased the fluorescence expression of eGFP compared with the group of PSB1C3-sulAp-eGFP. This result greatly demonstrates the excellent capability of Amp30E Amplification Device.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1891
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1101
    Illegal SapI.rc site found at 1734



Reference
[1] Wan, Xinyi, et al. "Cascaded amplifying circuits enable ultrasensitive cellular sensors for toxic metals." Nature chemical biology 15.5 (2019): 540-548.

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Parameters
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