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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. | + | <meta name="viewport" content="width=device-width, initial-scale=1.0"> |
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| − | <!-- Add more about the biology of this part here | + | <link rel="stylesheet" href="https://2022.igem.wiki/scut-china/static/css/part-public.css"> |
| − | ===Usage and Biology=== | + | </head> |
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| | + | <h2 style="font-weight: bold;">P<sub>ERG9</sub></h2> |
| | + | <h2>Introduction</h2> |
| | + | <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> |
| | + | <h2>Characterization</h2> |
| | + | <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> |
| | + | <img src="https://static.igem.wiki/teams/4263/wiki/parts/image/erg9e-min.jpg" alt=""> |
| | + | <h4>Figure 1 Gene circuit of P<em><sub>ERG9</sub>-EGFP-</em>terminator</h4> |
| | + | <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> |
| | + | <img src="https://static.igem.wiki/teams/4263/wiki/parts/image/perg9-min.jpg" alt=""> |
| | + | <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> |
| | + | <h2>Reference</h2> |
| | + | <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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| | <span class='h3bb'>Sequence and Features</span> | | <span class='h3bb'>Sequence and Features</span> |
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| | ===Functional Parameters=== | | ===Functional Parameters=== |
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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.
