Difference between revisions of "Part:BBa K385004:Experience"
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===Applications of BBa_K385004=== | ===Applications of BBa_K385004=== | ||
| − | 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 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). | 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). | ||
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[[Image:N25 + Gal-3.jpg|center|400 px]] | [[Image:N25 + Gal-3.jpg|center|400 px]] | ||
| − | A control culture of the same transformant was grown using glucose as the carbon source; these conditions do not activate the GAL promoter | + | 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. | Overall the results indicate that the N-peptide can be successfully expressed as a protein fusion with other standard parts. | ||
Latest revision as of 20:55, 24 October 2010
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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).
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.
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