Difference between revisions of "Part:BBa K3861017"

(Characterization)
Line 24: Line 24:
 
<html>
 
<html>
  
We chara
+
We wanted to know if the lactate-inducible lldR promoter is still functional in <i>Salmonella</i> minicells, as minicells only exhibit a limited lifespan and transcriptional and translational activity. To check this, we conducted a <a href="https://2021.igem.org/Team:Humboldt_Berlin/Experiments">lactate assay</a> with 2 replications using newly purified minicells following our optimized <a href="https://2021.igem.org/Team:Humboldt_Berlin/Contribution">minicell purification protocol</a>. 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).
  
 
<figure>
 
<figure>
 
<img src="https://2021.igem.org/wiki/images/8/8c/T--Humboldt_Berlin--plldR_characterisation.png" alt="PlldR-GFP" width="500">
 
<img src="https://2021.igem.org/wiki/images/8/8c/T--Humboldt_Berlin--plldR_characterisation.png" alt="PlldR-GFP" width="500">
<figcaption>Fig. 1 - Characterization of the lldR promoter using GFP as reporter. RFU = relative fluorescene unit; n = replications.</figcaption>
+
<figcaption>Fig. 1 - Characterization of the lldR promoter using GFP as reporter. RFU = relative fluorescene unit; n = number of replications.</figcaption>
 
</figure>
 
</figure>
 
</html>
 
</html>

Revision as of 20:11, 21 October 2021


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.