Difference between revisions of "Part:BBa K2213013"

Revision as of 00:36, 2 November 2017

araB_eutLK_LowPromoter_PduD(1-20)_mCherry_cgPPK2
araB_eutLK_LowPromoter_PduD(1-20)_mCherry_cgPPK2 is a composite of https://parts.igem.org/Part:BBa_K2213002, https://parts.igem.org/Part:BBa_K2213006 and https://parts.igem.org/Part:BBa_K2213003.

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.


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