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| | <h2 style="font-weight: bold;">P<sub>0547</sub></h2> | | <h2 style="font-weight: bold;">P<sub>0547</sub></h2> |
| | <h2>Introduction</h2> | | <h2>Introduction</h2> |
| − | <p>0547 promoter is a novol promising methanol inducible promoter in the yeast Pichia pastoris<sup>[1]</sup>.</p> | + | <p>0547 promoter is a novol promising methanol inducible promoter in the yeast <em>Pichia pastoris</em><sup>[1]</sup>.</p> |
| | <h2>Characterization</h2> | | <h2>Characterization</h2> |
| − | <p>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.</p> | + | <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/0547e.jpg" alt=""> | + | <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> | + | <h4>Figure 1 Gene circuit of P<em><sub>0547</sub>-EGFP</em>-terminator</h4> |
| − | <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 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.</p> | + | <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/p0547.jpg" alt=""> | + | <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> | + | <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>Reference</h2> | | <h2>Reference</h2> |
| − | <p>[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.</p> | + | <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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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.
