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<center> <h4>pSBC3-ADH1+gld+tGPD1 & pSBC3-PGK1+DAK+TPFK1<br><h4></center> | <center> <h4>pSBC3-ADH1+gld+tGPD1 & pSBC3-PGK1+DAK+TPFK1<br><h4></center> | ||
<h4>The enzyme GlyDH (gld) and DAK can be efficiently expressed in the yeast cell, and the gene of the | <h4>The enzyme GlyDH (gld) and DAK can be efficiently expressed in the yeast cell, and the gene of the | ||
− | + | enzyme GlyDH (gld) and DAK are integrated into the chromosome. In this pathway, GlyDH (Glycerol | |
− | + | dehydrogenase) can efficiently convert glycerol to DHA (1,3-Dihydroxyacetone), and then DAK | |
− | + | (Dihydroxyacetone kinase) phosphorylate DHA into DHAP, and then DHAP is catalyzed into acrylic acid | |
− | + | by ceaS2. | |
</h4> | </h4> | ||
<center> <img src="https://static.igem.org/mediawiki/2017/6/65/YCplac33-URA-gld-DAK.png" class="img-responsive"></center> | <center> <img src="https://static.igem.org/mediawiki/2017/6/65/YCplac33-URA-gld-DAK.png" class="img-responsive"></center> | ||
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<h4>In the yeast cells, the enzyme ceaS2 can be efficiently expressed and the gene of ceas2 is integrated into | <h4>In the yeast cells, the enzyme ceaS2 can be efficiently expressed and the gene of ceas2 is integrated into | ||
− | + | the chromosome. With the help of TPP (Thiamine pyrophosphate) and magnesium ions, ceaS2 can | |
− | + | catalyze the production of acrylic acid with DHAP (dihydroxy acetone phosphate) and G3P | |
− | + | (glyceraldehyde 3-phosphate ) as substrate. </h4><br> | |
</h4> | </h4> | ||
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<h4>4.pSBC3-TEF2+NOX+tRPS2<br><h4> | <h4>4.pSBC3-TEF2+NOX+tRPS2<br><h4> | ||
<h4>In this part, the promoter of NOX is TEF2 promoter, and the terminator is tRPS2 terminator. TEF2 promoter is | <h4>In this part, the promoter of NOX is TEF2 promoter, and the terminator is tRPS2 terminator. TEF2 promoter is | ||
− | + | a kind of promoter which has strong expression and tRPS2 terminator is a terminator with rather strong | |
− | + | expression ability. So they can both increase the expression of genes. At the same time they are constitutive | |
− | + | promoters and terminators, which have the ability to integrate gene fragment, NOX, into the chromosome | |
− | + | of yeast, thereby reducing the burden of plasmid expression in yeast. We linked this part with part | |
− | + | pSBC3-pTDH3+ceas2+tPFK1and the YCplac33 plasmid vector to form intact plasmid with LEU deficient. | |
</h4><br> | </h4><br> | ||
<h4> <div class="col-md-12" style="padding-top:30px"> | <h4> <div class="col-md-12" style="padding-top:30px"> | ||
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</div> | </div> | ||
<h4>It can efficiently express the enzyme NOX in yeast and integrate NOX into the chromosome. Since GlyDH | <h4>It can efficiently express the enzyme NOX in yeast and integrate NOX into the chromosome. Since GlyDH | ||
− | + | is an NAD+ -dependent enzyme, NOX and CAT(which already exists in yeast) provide the required | |
− | + | reduction force for GLYDH through the two layers of substrate level cycle. | |
</h4><br> | </h4><br> | ||
</h4> | </h4> |
Latest revision as of 02:33, 2 November 2017