Revision as of 05:42, 27 September 2022

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

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