Difference between revisions of "Part:BBa K3384314"

 
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<partinfo>BBa_K3384314 short</partinfo>
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Multiple copies of putative Ste12 binding sites are prevalent in pheromone-responsive promoters of Saccharomyces cerevisiae, which are also known as pheromone response elements (PREs). pprm1 contains 3 × PREs which is in the opposite direction of the promoter. pprm1 Ultra is a modified pheromone-responsive promoter. It  contains 6 × PREs in the opposite direction of the promoter.
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[[File:NJTech_China_Ste5ΔN-CTM-1.png|width='100%' valign='top'| |center|thumb|550px|''<b>Fig.1</b>Structure and function of Ste5ΔN-CTM.]]
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<br>
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===Characterization===
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When assembled with promotor BBa_K3384314, BBa_K3112009, and BBa_K3384311 in pRS415, this construct expressed GFP. Then the activity of these promoters can be quantitatively measured through flow cytometer. As is shown in figure 1, the fluorescence intensity of the pprm1 is higher than that of the pprm1 Ultra under high concentration treatment conditions of pheromones.
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The pheromone concentration has no significant effect on the GFP expression intensity under the control of pprm1 Ultra, indicating that this engineered pprm1 remains a stable expression level when induced by pheromone. As pprm1 Ultra can provide a stable expression level regardless of the fluctuations in external conditions, it is expected to be applied to gene expression regulation in cell factory.
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<p> </p>
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[[File:NJTech_China_Ste5ΔN-CTM-2.png|width='100%' valign='top'| |center|thumb|550px|''<b>Fig.1</b>The growth of the Ste5ΔN-CTM strain and BY4741 wild-type strain on the plate with galactosyl as the sole carbon source.]]
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<p> </p>
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[[File:NJTech_China_Ste5ΔN-CTM-3.png|width='100%' valign='top'| |center|thumb|550px|''<b>Fig.2</b>The growth of the Ste5ΔN-CTM strain and BY4741 wild-type strain on the plate with glucose as the sole carbon source.]]
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===Reference===
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1. LSengupta, P., and Cochran, B. H. (1990) The Pre and Pq Box Are Functionally Distinct Yeast Pheromone Response Elements, Molecular and Cellular Biology 10, 6809-6812.
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3384314 SequenceAndFeatures</partinfo>
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===Functional Parameters===
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<partinfo>BBa_K3384314 parameters</partinfo>
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Revision as of 03:15, 27 October 2020


pprm1 Ultra

Multiple copies of putative Ste12 binding sites are prevalent in pheromone-responsive promoters of Saccharomyces cerevisiae, which are also known as pheromone response elements (PREs). pprm1 contains 3 × PREs which is in the opposite direction of the promoter. pprm1 Ultra is a modified pheromone-responsive promoter. It contains 6 × PREs in the opposite direction of the promoter.


Fig.1Structure and function of Ste5ΔN-CTM.


Characterization

When assembled with promotor BBa_K3384314, BBa_K3112009, and BBa_K3384311 in pRS415, this construct expressed GFP. Then the activity of these promoters can be quantitatively measured through flow cytometer. As is shown in figure 1, the fluorescence intensity of the pprm1 is higher than that of the pprm1 Ultra under high concentration treatment conditions of pheromones. The pheromone concentration has no significant effect on the GFP expression intensity under the control of pprm1 Ultra, indicating that this engineered pprm1 remains a stable expression level when induced by pheromone. As pprm1 Ultra can provide a stable expression level regardless of the fluctuations in external conditions, it is expected to be applied to gene expression regulation in cell factory.



Fig.1The growth of the Ste5ΔN-CTM strain and BY4741 wild-type strain on the plate with galactosyl as the sole carbon source.



Fig.2The growth of the Ste5ΔN-CTM strain and BY4741 wild-type strain on the plate with glucose as the sole carbon source.



Reference

1. LSengupta, P., and Cochran, B. H. (1990) The Pre and Pq Box Are Functionally Distinct Yeast Pheromone Response Elements, Molecular and Cellular Biology 10, 6809-6812.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]