Difference between revisions of "Part:BBa K385003:Experience"
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===Applications of BBa_K385003=== | ===Applications of BBa_K385003=== | ||
| − | The N-peptide 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 in the single copy shuttle vector pRS415. | + | The N-peptide 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). | 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). | ||
Revision as of 11:02, 24 September 2010
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Applications of BBa_K385003
The N-peptide 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).
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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