Part:BBa_K562012:Experience

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

User Reviews

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••••• iGEM Dundee 2011

This part was seen to work in practice. The PduD20 tag fused to GFP resulted in fluorescent cells (Figure 1A) and, when co-expressed in the presence of the microcompartment encoded by BBa_K562009 was found to lead to distinct, foci (Figure 1B) suggesting targeting to - or into -the microcompartment.

Results

E. coli was transformed with BBa_K562012 alone or in combination with BBa_K562009 and grown overnight in LB medium under aerobic conditions. A sample of whole cells was taken, washed in phosphate-buffered saline, and applied to a microscope slide using CellTak adhesive. The cells were then observed by confocal microscopy and shown to be fluorescent (Figure 1).

Strikingly, co-expression of the microcompartment led to clustering of GFP in spherical bodies, suggesting the GFP was being targeted into the microcompartment. The cells were also observed to be elongated although NO GROWTH DEFECTS were detected.

For futher characterisation of the Pdu20-GFP - Microcompartment interaction please see the experience page of BBa_K562009.

Figure 1: (A) PduD20-GFP expressed alone in E. coli. (B) PduD20-GFP co-expressed with a bacterial microcompartment encoded by BBa_K562009. .

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