Difference between revisions of "Part:BBa K4263002"

 
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    <h2 style="font-weight: bold;">P<sub>DAS2</sub></h2>
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    <h2>Introduction</h2>
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    <p>DAS2 is a promoter derived from dedihydroxy-acetone synthase (<em>DAS</em>) gene<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>DAS2</sub></em>, we construct "P<em><sub>DAS2</sub>-EGFP-</em>terminator" (Figure 1). If P<em><sub>DAS2</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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    <img src="https://static.igem.wiki/teams/4263/wiki/parts/image/das2e-min.jpg" alt="">
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    <h4>Figure 1 Gene circuit of P<em><sub>DAS2</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. 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/pdas2-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] Vogl T, Kickenweiz T, Pitzer J, et al. Engineered bidirectional promoters enable rapid multi-gene co-expression optimization[J]. Nat Commun, 2018,9(1):3589.</p>
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Revision as of 15:37, 26 September 2022

K4263002

PDAS2

Introduction

DAS2 is a promoter derived from dedihydroxy-acetone synthase (DAS) gene[1].

Characterization

In order to test the function of PDAS2, we construct "PDAS2-EGFP-terminator" (Figure 1). If PDAS2 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 PDAS2-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. 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] Vogl T, Kickenweiz T, Pitzer J, et al. Engineered bidirectional promoters enable rapid multi-gene co-expression optimization[J]. Nat Commun, 2018,9(1):3589.