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<h2 class="editContent">Model</h2> | <h2 class="editContent">Model</h2> | ||
− | <p>We designed our constructs (pGMP1- | + | <p>We designed our constructs (<em>pGMP1</em>-<em>P<sub>AOX1</sub></em>-GFP, <em>pAPM1</em>-<em>P<sub>AOX1</sub></em>-GFP and <em>pGMP1</em>-<em>pAPM1</em>-<em>P<sub>AOX1</sub></em>-GFP) using homogenous promoters and transcription factors in <em>P. pastoris GS115</em> to moderately up-regulate the activity of the <em>AOX1</em> promoter. To get an idea of how much more efficient our constructs will be compared with the wild-type <em>P. pastoris</em>, we modeled our constructs based on various statistics published in Wang et al., 2016, Shi et al., 2019 and Liang et al., 2012. We then calculated basic parameters for each promoter and transcription factor in our three constructs, specifically, the activity of a promoter under the effect of 1 unit transcription factor. As it turned out, we relatively successfully calculated the eventual GFP expression by <em>P<sub>AOX1</sub></em> in each of our constructs and compared that to the GFP expression by <em>P<sub>AOX1</sub></em> in the <em>P<sub>AOX1</sub></em>-GFP strain (GFP expressed by <em>AOX1</em> promoter homologously reintegrated into wild type <em>P. pastoris GS115</em>, in which GFP indicates <em>P<sub>AOX1</sub></em> activity).</p> |
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Revision as of 15:22, 19 October 2019