Composite

Part:BBa_K2824004

Designed by: Jiang Qiaochu   Group: iGEM18_NEU_China_B   (2018-10-05)


lldPRD operon promoter-GFP

Long Description

LLDPRD can sense the presence of lactic acid, in the absence of lactic acid, only a lower background level of expression, but in the presence of lactic acid can open expression, gene transcription. We added the gfp gene to the gene, enabling the part to identify the presence of lactic acid using green fluorescence as a marker.

Characterization

The part is one of the most critical components in the system to detect whether the system correctly perceives the presence of lactic acid and triggers the corresponding gene expression. In our experiment, when there was no lactic acid, the content of GFP was very low and the fluorescence intensity was weak, which was slightly higher than that of the blank control group. When lactic acid exists, the content of GFP changes with the increase of lactic acid concentration. The fluorescence intensity increases first and then decreases, and the expression level is the highest under the condition of lactic acid concentration about 1mMol/L. Then we do a group of more detailed data and make real time measurement using optical fiber within 10 minutes. The results show that the fluorescence signal does not rise in about a minute, which shows that the system running time of this project is relatively short. This is conducive to real-time detection in production.

The following diagram shows the fluorescence intensity curve of lactic acid concentration of 1 mM and induction of 5 minutes (Figure 1).

T--NEU_China_B--p700.png

Figure 1:lldPRD operon promoter-GFP 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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 335
    Illegal BsaI.rc site found at 994


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