Difference between revisions of "Part:BBa K4263004"

 
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Major ADP/ATP carrier of the mitochondrial inner membrane
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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>PET9</sub></h2>
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    <h2>Introduction</h2>
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    <p>PET9 promoter is the major ADP/ATP carrier of the mitochondrial inner membrane of <em>Pichia pastoris</em><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>PET9</sub></em>, we construct "P<em><sub>PET9</sub>-EGFP-</em>terminator" (Figure 1). If P<em><sub>PET9</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>PET9</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, which is in line with literature description<sup>[1]</sup>. 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/ppet9-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 15:39, 26 September 2022

K4263004

PPET9

Introduction

PET9 promoter is the major ADP/ATP carrier of the mitochondrial inner membrane of Pichia pastoris[1].

Characterization

In order to test the function of PPET9, we construct "PPET9-EGFP-terminator" (Figure 1). If PPET9 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 PPET9-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, 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] 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.