Coding

Part:BBa_K385004:Experience

Designed by: Krystal Annand, Joseph Hoare, Stephen Lam, Justyna Kucia   Group: iGEM10_Aberdeen_Scotland   (2010-09-27)
Revision as of 12:04, 27 September 2010 by I.stansfield (Talk | contribs) (Applications of BBa_K385004)

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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. The results (Figure 2) show no GFP fluorescence.

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

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