Part:BBa_K562013: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

UNIQ65ea7b2bc9a6a56d-partinfo-00000000-QINU

••• iGEM Dundee 2011

This part was seen to work in practice. The PduD40 tag fused to GFP resulted in fluorescent cells (Figure 1). Co-expression of a bacterial microcompartment BBa_K562009 led to the development of bright foci (Figure 1) suggesting GFP was being compartmentalised.

Results

E. coli was transformed with BBa_K562013 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).

Co-expression of the microcompartment led to foci of GFP in spherical bodies, suggesting the PduD40-GFP-HA fusion protein was being targeted into the microcompartment.

Figure 1: PduD40-GFP expressed in E. coli with a bacterial microcompartment encoded by BBa_K562009.

UNIQ65ea7b2bc9a6a56d-partinfo-00000006-QINU