Difference between revisions of "Part:BBa K2213013"
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<b>Figure 1.</b> Fluorescence microscopy images of promoter-PduD associated mCherry and DAPI stained polyphosphate.<br>
 
<b>Figure 1.</b> Fluorescence microscopy images of promoter-PduD associated mCherry and DAPI stained polyphosphate.<br>
 
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As shown, there is a heterogeneous distribution of fluorescence within the cells for both signals and they are approximately in the same areas. This heterogeneous distribution of DAPI indicates the presence of polyphosphate and proves the activity of our PPK along with its successful localization into our bacterial microcompartment. The localisation can be determined using the physical location of both fluorescence signals within the cell.<br>
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As shown, there is a heterogeneous distribution of fluorescence within the cells for both signals and they are approximately in the same areas. This heterogeneous distribution of DAPI indicates the presence of polyphosphate and proves the activity of the PPK along with its successful localization into the Eut microcompartment. The localisation can be determined using the physical location of both fluorescence signals within the cell. These findings help establish a proof-of-concept functionality of the system and demonstrate successful Eut subunit expression, successful tag localisation and successful PPK activity.<br>
 
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Revision as of 22:43, 1 November 2017


araB_eutLK_LowPromoter_PduD(1-20)_mCherry_cgPPK2

Characterisation

This part was expressed alongside EutSMN (https://parts.igem.org/Part:BBa_K2213012) to form EutSMNLK+Low_PduD+PPK which was characterized as follows.

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

LowSMNLKDAPIcells.png
Figure 1. Fluorescence microscopy images of promoter-PduD associated mCherry and DAPI stained polyphosphate.

As shown, there is a heterogeneous distribution of fluorescence within the cells for both signals and they are approximately in the same areas. This heterogeneous distribution of DAPI indicates the presence of polyphosphate and proves the activity of the PPK along with its successful localization into the Eut microcompartment. The localisation can be determined using the physical location of both fluorescence signals within the cell. These findings help establish a proof-of-concept functionality of the system and demonstrate successful Eut subunit expression, successful tag localisation and successful PPK activity.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1300
    Illegal NheI site found at 2701
    Illegal NheI site found at 2724
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1239
    Illegal BamHI site found at 2164
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1074
    Illegal AgeI site found at 1775
    Illegal AgeI site found at 1952
    Illegal AgeI site found at 2252
    Illegal AgeI site found at 2315
    Illegal AgeI site found at 2362
    Illegal AgeI site found at 3601
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2839
    Illegal SapI site found at 1056
    Illegal SapI.rc site found at 4280