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     <h2>(2) Characterization</h2>
 
     <h2>(2) 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>
 
     <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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     <h4>Figure 1 Gene circuit of P<sub>0547</sub>-EGFP</h4>
 
     <h4>Figure 1 Gene circuit of P<sub>0547</sub>-EGFP</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>
 
     <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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     <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 EGFP gene.</h4>
 
     <h2>(3) Reference</h2>
 
     <h2>(3) Reference</h2>

Revision as of 08:38, 26 September 2022

Document

P0547

(1) Introduction

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

(2) 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

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.

(3) 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.