Part:BBa_K3861017

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).

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