Difference between revisions of "Part:BBa K4263007"

 
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AACATCCAAAGACGAAAGGTTGAATGAAACCTTTTTGCCATCCGACATCCACAGGTCCATTCTCACACATAAGTGCCAAACGCAACAGGAGGGGATACACTAGCAGCAGACCGTTGCAAACGCAGGACCTCCACTCCTCTTCTCCTCAACACCCACTTTTGCCATCGAAAAACCAGCCCAGTTATTGGGCTTGATTGGAGCTCGCTCATTCCAATTCCTTCTATTAGGCTACTAACACCATGACTTTATTAGCCTGTCTATCCTGGCCCCCCTGGCGAGGTTCATGTTTGTTTATTTCCGAATGCAACAAGCTCCGCATTACACCCGAACATCACTCCAGATGAGGGCTTTCTGAGTGTGGGGTCAAATAGTTTCATGTTCCCCAAATGGCCCAAAACTGACAGTTTAAACGCTGTCTTGGAACCTAATATGACAAAAGCGTGATCTCATCCAAGATGAACTAAGTTTGGTTCGTTGAAATGCTAACGGCCAGTTGGTCAAAAAGAAACTTCCAAAAGTCGGCATACCGTTTGTCTTGTTTGGTATTGATTGACGAATGCTCAAAAATAATCTCATTAATGCTTAGCGCAGTCTCTCTATCGCTTCTGAACCCCGGTGCACCTGTGCCGAAACGCAAATGGGGAAACACCCGCTTTTTGGATGATTATGCATTGTCTCCACATTGTATGCTTCCAAGATTCTGGTGGGAATACTGCTGATAGCCTAACGTTCATGATCAAAATTTAACTGTTCTAACCCCTACTTGACAGCAATATATAAACAGAAGGAAGCTGCCCTGTCTTAAACCTTTTTTTTTATCATCATTATTAGCTTACTTTCATAATTGCGACTGGTTCCAATTGACAAGCTTTTGATTTTAACGACTTTTAACGACAACTTGAGAAGATCAAAAAACAACTAATTATTCGAAACG
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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>AOX1</sub></h2>
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    <h2>Introduction</h2>
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    <p>AOX1 promoter is a key promoter in the methylotrophic yeast <em>Pichia pastoris</em>.</p>
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    <h2>Characterization</h2>
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    <p>In order to test the function of P<em><sub>AOX1</sub></em>, we construct "P<em><sub>AOX1</sub>-EGFP</em>-terminator" (Figure 1). If P<em><sub>AOX1</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>AOX1</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 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>
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    <img src="https://static.igem.wiki/teams/4263/wiki/parts/image/paox1-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] Xuan Y, Zhou X, Zhang W, et al. An upstream activation sequence controls the expression of <em>AOX1</em> gene in Pichia pastoris[J]. FEMS Yeast Res, 2009,9(8):1271-1282.</p>
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Revision as of 15:42, 26 September 2022

K4263007

PAOX1

Introduction

AOX1 promoter is a key promoter in the methylotrophic yeast Pichia pastoris.

Characterization

In order to test the function of PAOX1, we construct "PAOX1-EGFP-terminator" (Figure 1). If PAOX1 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 PAOX1-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] Xuan Y, Zhou X, Zhang W, et al. An upstream activation sequence controls the expression of AOX1 gene in Pichia pastoris[J]. FEMS Yeast Res, 2009,9(8):1271-1282.