Regulatory

Part:BBa_K822000

Designed by: Jarle Pahr   Group: iGEM12_NTNU_Trondheim   (2012-09-23)

lldPRD operon promoter + RBS from E. coli

This is the promoter region for the lldPRD operon (previously named lct) of E. coli. The lldPRD operon encodes a lactate-dehydrogenase, and the lldPRD promoter has been shown to be induced by L-lactate.

The promoter is repressed/activated by the regulatory protein lldR encoded by the operon. In the absence of L-lactate, lldR binds to two operator sites O1 and O2 in the promoter region and inhibits expression. [1]

The activity of the promoter may be repressed by chloramphenicol and nutrient-rich growth environment. [2]

Characterization by ETH iGEM 2015

Check the characterization done by ETH Zurich 2015 iGEM team in the experience page.

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



References

[1]: Dual role of lldR in regulation of the lldPRD operon, involved in L-lactate metabolism in E. coli. Aguilera et al. J. Bacteriol. 2008, 190(8):2997

[2]: Inducible Membrane-bound L-Lactate Dehydrogenase from Escherichia coli - purification and properties. Futai, M. and Kimura, H. Journal of Biological Chemistry. 1977, 252(16):5820-5827.


Improvement by NEU-CHINA-B iGEM 2018

BBa_k2824006:T7-lldPRD operon promoter-GFP>/h2> <h2> Improve the Characterization of BBa_k82000

In the following figure, we compared the expression of lldPRD operon promoter-GFP and T7-lldPRD operon promoter-GFP. The T7 promoter could enhance gene expression when the concentration of lactic acid was less than 1 m mol/L. Considering that the lactate concentration in yogurt is generally no more than 1m mol/L, the conclusion can given that the T7 promoter can enhance the signal intensity for our experiments (Figure 1). <p> Figure 1:lldPRD operon promoter-GFP vs T7-lldPRD operon promoter-GFP

To see more details about the construction and result, click the hyperlink below: T7-lldPRD operon promoter-GFP composite:BBa_k2824006(http://parts.igem.org/Part:BBa_K2824006).

BBa_k2824008:Lldr-T7-lldPRD operon promoter-GFP

Improve the Characterization of BBa_k82000

<p> We can easily find that the Lldr-T7-lldPRD operon promoter-GFP part are in a higher level of expression comparing to the lldPRD operon promoter-GFP when the lactate concentration is lower than nearly 1.5 mM. Considering that the lactate concentration in yogurt is generally no more than 1 mM, the conclusion can give that the T7 promoter can enhance the signal intensity for our experiments (Figure 2).

T--NEU_China_B--8end0.png

Figure 1:Lldr-T7-lldPRD operon promoter-GFP

T--NEU_China_B--8end1.png

Figure 2:lldPRD operon promoter-GFP vs Lldr-T7-lldPRD operon promoter-GFP

To see more details about the construction and result, click the hyperlink below:

Lldr-T7-lldPRD operon promoter-GFP composite: BBa_k2824008(http://parts.igem.org/Part:BBa_K2824008)
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