Coding

Part:BBa_K385004:Experience

Designed by: Krystal Annand, Joseph Hoare, Stephen Lam, Justyna Kucia   Group: iGEM10_Aberdeen_Scotland   (2010-09-27)

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.

Applications of BBa_K385004

The N-peptide tandem repeat reading frame was fused in-frame to GFP to make a translational fusion. It was placed under control of the yeast GAL1 promoter (BBa_J63006), and transformed into yeast Saccharomyces cerevisiae in the single copy shuttle vector pRS415.

The transformants were grown overnight in synthetic defined medium containing 2% w/v galactose, and observed using a fluorescence microscope optimised for GFP visualisation (Figure 1).

N25 + Gal-3.jpg

A control culture of the same transformant was grown using glucose as the carbon source; these conditions do not activate the GAL promoter, and as expected no GFP fluorescence was detectable (data not shown).

Overall the results indicate that the N-peptide can be successfully expressed as a protein fusion with other standard parts.

User Reviews

UNIQb6dd7d3a4afd3e7a-partinfo-00000000-QINU UNIQb6dd7d3a4afd3e7a-partinfo-00000001-QINU