Difference between revisions of "Part:BBa K4263003"

 
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Phosphoglycerate kinase 1
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
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    <h2 style="font-weight: bold;">P<sub>PGK1</sub></h2>
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    <h2>Introduction</h2>
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    <p>PGK1 promoter is the promoter for the glycolytic enzyme 3-phosphoglycerate kinase<sup>[1]</sup>.</p>
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    <h2>Characterization</h2>
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    <p>In order to test the function of P<em><sub>PGK1</sub></em>, we construct "P<em><sub>PGK1</sub>-EGFP-</em>terminator" (Figure 1). If P<em><sub>PGK1</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>
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===Functional Parameters===
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    <h4>Figure 1 Gene circuit of P<em><sub>PGK1</sub>-EGFP-</em>terminator</h4>
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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 decreased during 24-72h. At the same time, we measured the growth curve of the strains.</p>
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    <img src="https://static.igem.wiki/teams/4263/wiki/parts/image/ppgk1-min.jpg" alt="">
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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>
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    <h2>Reference</h2>
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    <p>[1] Stadlmayr G, Mecklenbrauker A, Rothmuller M, et al. Identification and characterisation of novel <em>Pichia pastoris</em> promoters for heterologous protein production[J]. J Biotechnol, 2010,150(4):519-529.</p>
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Revision as of 15:38, 26 September 2022

K4263003

PPGK1

Introduction

PGK1 promoter is the promoter for the glycolytic enzyme 3-phosphoglycerate kinase[1].

Characterization

In order to test the function of PPGK1, we construct "PPGK1-EGFP-terminator" (Figure 1). If PPGK1 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 PPGK1-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 decreased during 24-72h. 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] Stadlmayr G, Mecklenbrauker A, Rothmuller M, et al. Identification and characterisation of novel Pichia pastoris promoters for heterologous protein production[J]. J Biotechnol, 2010,150(4):519-529.