Composite

Part:BBa_K3861017

Designed by: Mario Delgadillo   Group: iGEM21_Humboldt_Berlin   (2021-10-01)
Revision as of 21:50, 21 October 2021 by Martje1998 (Talk | contribs)


PlldR-GFP

Introduction

Composite part containing the PlldR promoter (BBa_K1847008) fused to GFP (BBa_E0840) as a reporter gene. Both parts were already characterized in depth by other teams. We used this part in Salmonella Typhimurium.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 78
    Illegal NheI site found at 101
  • 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 816


Characterization

We wanted to know if the lactate-inducible lldR promoter is still functional in Salmonella minicells, as minicells only exhibit a limited lifespan and transcriptional and translational activity. To check this, we conducted a lactate assay with 2 replications using newly purified minicells following our optimized minicell purification protocol. The GFP expression is increasing with rising lactate-concentration, thus we concluded that minicells have enough expression capacity available for GFP expression and that the lldR promoter is functional (Fig. 1).

PlldR-GFP
Fig. 1 - Characterization of the lldR promoter using GFP as reporter. RFU = relative fluorescene unit; n = number of replications.

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