Difference between revisions of "Part:BBa K3384312"
 
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p<em>fus2</em> is a pheromone-responsive promoter in <em>Saccharomyces cerevisiae</em>. p<em>fus2</em> contains 2 copies of pheromone responsive elment (PRE), one of which is in the opposite direction of the promoter and the other one is in the same direction of the promoter.
 
p<em>fus2</em> is a pheromone-responsive promoter in <em>Saccharomyces cerevisiae</em>. p<em>fus2</em> contains 2 copies of pheromone responsive elment (PRE), one of which is in the opposite direction of the promoter and the other one is in the same direction of the promoter.
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===Characterization===
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When assembled with GFP <partinfo>BBa_K3112009</partinfo>, and CYC1 terminator <partinfo>BBa_K3384311</partinfo> in pRS415, p<em>fus2</em> regulated the expression of GFP. After pheromone induction, the intensity of p<em>fus2</em> is reflected by the fluorescence intensity of GFP which is quantified by flow cytometry.
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When OD=0.6, α-factor was added for induction, the results showed that the pheromone has a significant inducing effect on p<em>fus2</em>. The results measured by flow cytometry is shown in figure 1. Within a certain range, which is 1-25μM, the higher the concentration of pheromone, the more obvious the induction effect.
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[[File:NJTech_China_pfus2.png|width='100%' valign='top'| |center|thumb|550px|''<b>Fig.1</b> The fluorescence intensity of GFP expressed by pfus2 induced by different concentrations of pheromone.]]
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===Reference===
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Elion, E. A., Trueheart, J., and Fink, G. R. (1995) Fus2 Localizes near the Site of Cell-Fusion and Is Required for Both Cell-Fusion and Nuclear Alignment during Zygote Formation, Journal of Cell Biology 130, 1283-1296.

Latest revision as of 07:55, 27 October 2020

pfus2

pfus2 is a pheromone-responsive promoter in Saccharomyces cerevisiae. pfus2 contains 2 copies of pheromone responsive elment (PRE), one of which is in the opposite direction of the promoter and the other one is in the same direction of the promoter.


Characterization

When assembled with GFP BBa_K3112009, and CYC1 terminator BBa_K3384311 in pRS415, pfus2 regulated the expression of GFP. After pheromone induction, the intensity of pfus2 is reflected by the fluorescence intensity of GFP which is quantified by flow cytometry. When OD=0.6, α-factor was added for induction, the results showed that the pheromone has a significant inducing effect on pfus2. The results measured by flow cytometry is shown in figure 1. Within a certain range, which is 1-25μM, the higher the concentration of pheromone, the more obvious the induction effect.

Fig.1 The fluorescence intensity of GFP expressed by pfus2 induced by different concentrations of pheromone.


Reference

Elion, E. A., Trueheart, J., and Fink, G. R. (1995) Fus2 Localizes near the Site of Cell-Fusion and Is Required for Both Cell-Fusion and Nuclear Alignment during Zygote Formation, Journal of Cell Biology 130, 1283-1296.