Difference between revisions of "Part:BBa K4263000"

 
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     <h1>P<sub>0547</sub></h1>
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     <h2 style="font-weight: bold;">P<sub>0547</sub></h2>
     <h2>(1) Introduction</h2>
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     <h2>Introduction</h2>
     <h3>0547 promoter is a novol promising methanol inducible promoter in the yeast Pichia pastoris<sup>[1]</sup>.</h3>
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     <p>0547 promoter is a novol promising methanol inducible promoter in the yeast <em>Pichia pastoris</em><sup>[1]</sup>.</p>
     <h2>(2) Characterization</h2>
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     <h2>Characterization</h2>
     <h3>In order to test the function of P<sub>0547</sub>, we construct "P<sub>0547</sub>-EGFP- terminator"(Figure 1). If P<sub>0547</sub> is functional, we can test the fluorescence intensity of EGFP in supernatant samples obtained from the culture of recombinant <em>P.pastoris</em> GS115 strain.</h3>
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     <p>In order to test the function of P<em><sub>0547</sub></em>, we construct "P<em><sub>0547</sub>-EGFP</em>-terminator" (Figure 1). If P<em><sub>0547</sub></em> is functional, we can test the fluorescence intensity of EGFP in supernatant samples obtained from the culture of recombinant <em>P.pastoris</em> GS115 strain.</p>
     <img src="https://static.igem.wiki/teams/4263/wiki/parts/p0547.jpg" alt="">
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     <img src="https://static.igem.wiki/teams/4263/wiki/parts/image/0547e-min.jpg" alt="">
     <h4>Figure 1 Gene circuit of P<sub>0547</sub>-EGFP</h4>
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     <h4>Figure 1 Gene circuit of P<em><sub>0547</sub>-EGFP</em>-terminator</h4>
     <h3>Our results matched the general expected trend (Figure 2). After fermentation experiment in BMMY medium containing 1% methanol. The fluorescence intensity of the samples of recombinant <em>P.pastoris</em> GS115 containing the EGFP gene gradually increased over time, which is in line with literature description<sup>[1]</sup>. At the same time, we measured the growth curve of the strains.</h3>
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     <p>Our results matched the general expected trend (Figure 2). After fermentation experiment in BMMY medium containing 1% methanol. The fluorescence intensity of the samples of recombinant <em>P.pastoris</em> GS115 containing the <em>EGFP</em> gene gradually increased over time, which is in line with literature description<sup>[1]</sup>. At the same time, we measured the growth curve of the strains.</p>
     <img src="https://static.igem.wiki/teams/4263/wiki/parts/0547e.jpg" alt="">
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     <img src="https://static.igem.wiki/teams/4263/wiki/parts/image/p0547-min.jpg" alt="">
     <h4>Figure 2 Fluorescence intensity and OD600 absorbance of samples obtained at different time points from the culture of corresponding recombinant <em>P.pastoris</em> GS115 containing EGFP gene.</h4>
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     <h4>Figure 2 Fluorescence intensity and OD600 absorbance of samples obtained at different time points from the culture of corresponding recombinant <em>P.pastoris</em> GS115 containing <em>EGFP</em> gene.</h4>
     <h2>(3) Reference</h2>
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     <h2>Reference</h2>
     <h3>[1] Xu N, Zhu J, Zhu Q, et al. Identification and characterization of novel promoters for recombinant protein production in yeast Pichia pastoris[J]. Yeast, 2018,35(5):379-385.</h3>
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     <p>[1] Xu N, Zhu J, Zhu Q, et al. Identification and characterization of novel promoters for recombinant protein production in yeast <em>Pichia pastoris</em>[J]. Yeast, 2018,35(5):379-385.</p>
 
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K4263000 SequenceAndFeatures</partinfo>
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<!-- Uncomment this to enable Functional Parameter display
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===Functional Parameters===
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<partinfo>BBa_K4263000 parameters</partinfo>

Latest revision as of 16:05, 30 September 2022

K4263000

P0547

Introduction

0547 promoter is a novol promising methanol inducible promoter in the yeast Pichia pastoris[1].

Characterization

In order to test the function of P0547, we construct "P0547-EGFP-terminator" (Figure 1). If P0547 is functional, we can test the fluorescence intensity of EGFP in supernatant samples obtained from the culture of recombinant P.pastoris GS115 strain.

Figure 1 Gene circuit of P0547-EGFP-terminator

Our results matched the general expected trend (Figure 2). After fermentation experiment in BMMY medium containing 1% methanol. The fluorescence intensity of the samples of recombinant P.pastoris GS115 containing the EGFP gene gradually increased over time, which is in line with literature description[1]. At the same time, we measured the growth curve of the strains.

Figure 2 Fluorescence intensity and OD600 absorbance of samples obtained at different time points from the culture of corresponding recombinant P.pastoris GS115 containing EGFP gene.

Reference

[1] Xu N, Zhu J, Zhu Q, et al. Identification and characterization of novel promoters for recombinant protein production in yeast Pichia pastoris[J]. Yeast, 2018,35(5):379-385.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 416
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 416
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 416
  • 1000
    COMPATIBLE WITH RFC[1000]