Part:BBa_K2213012

LacUV5_EutS_Tet_EutMN
LacUV5_EutS_Tet_EutMN is a composite of parts https://parts.igem.org/Part:BBa_K2213000 and https://parts.igem.org/Part:BBa_K2213001.

Figure 1: Schematic overview of LacUV5_EutS_Tet_EutMN (BBa_K2213012)

Characterisation

This part was co-transformed with EutLK-Low-PduD-mCherry-PPK (https://parts.igem.org/Part:BBa_K2213013) to form EutSMNLK+Low_PduD+PPK which was characterized as follows.

A 24 hour induction was DAPI stained to determine microcompartment formation and tag localisation.


Figure 2: Fluorescence microscopy images of promoter-PduD associated mCherry and DAPI stained polyphosphate.



Figure 3. Visible light, mCherry and DAPI signals from DAPI stained E. coli, lacking PPK.

DAPI staining and mCherry fluorescence within E.coli appear to colocalised, and locally enriched to foci at particular sub-cellular sites (Figure 2). This heterogeneous distribution of mCherry is induced by the presence of promoter-PduD-mCherry-PPK, with controls lacking the PPK element showing largely ubiquitous mCherry fluorescence throughout the cell (Figure 3). We conclude these results to indicate successful tag localisation and by proxy successful expression of Eut subunits.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 1260
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 4125
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 3451