Difference between revisions of "Part:BBa K4263005"

 
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Squalene synthase (SS) is the key precursor and first committed enzyme of the sterol biosynthesis pathway.ERG9 gene that encodes for SS.
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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>ERG9</sub></h2>
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    <h2>Introduction</h2>
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    <p>Squalene synthase (SS) is the key precursor and first committed enzyme of the sterol biosynthesis pathway. ERG9 gene that encodes for SS<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>ERG9</sub></em>, we construct "P<em><sub>ERG9</sub>-EGFP-</em>terminator" (Figure 1). If P<em><sub>ERG9</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>ERG9</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 was essentially unchanged 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/perg9-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] Lee P Y, Yong V C, Rosli R, et al. Cloning, expression and purification of squalene synthase from Candida tropicalis in <em>Pichia pastoris</em>[J]. Protein Expr Purif, 2014,94:15-21.</p>
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Revision as of 15:40, 26 September 2022

K4263005

PERG9

Introduction

Squalene synthase (SS) is the key precursor and first committed enzyme of the sterol biosynthesis pathway. ERG9 gene that encodes for SS[1].

Characterization

In order to test the function of PERG9, we construct "PERG9-EGFP-terminator" (Figure 1). If PERG9 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 PERG9-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 was essentially unchanged 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] Lee P Y, Yong V C, Rosli R, et al. Cloning, expression and purification of squalene synthase from Candida tropicalis in Pichia pastoris[J]. Protein Expr Purif, 2014,94:15-21.