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<td><a target="_blank" href="http://parts.igem.org/Part:BBa_K2347005">BBa_K2347005</a></td> | <td><a target="_blank" href="http://parts.igem.org/Part:BBa_K2347005">BBa_K2347005</a></td> | ||
<td>composite</td> | <td>composite</td> | ||
− | <td> | + | <td>DHAP/G3P->acrylic acid</td> |
<td>2862bp</td> | <td>2862bp</td> | ||
</tr> | </tr> | ||
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<td><a target="_blank" href="http://parts.igem.org/Part:BBa_K2347006">BBa_K2347006</a></td> | <td><a target="_blank" href="http://parts.igem.org/Part:BBa_K2347006">BBa_K2347006</a></td> | ||
<td>composite</td> | <td>composite</td> | ||
− | <td> | + | <td>glycerol->DHA</td> |
<td>2862bp</td> | <td>2862bp</td> | ||
</tr> | </tr> | ||
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<td><a target="_blank" href="http://parts.igem.org/Part:BBa_K2347007">BBa_K2347007</a></td> | <td><a target="_blank" href="http://parts.igem.org/Part:BBa_K2347007">BBa_K2347007</a></td> | ||
<td>composite</td> | <td>composite</td> | ||
− | <td> | + | <td>DHA->DHAP</td> |
<td>2862bp</td> | <td>2862bp</td> | ||
</tr> | </tr> | ||
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<td><a target="_blank" href="http://parts.igem.org/Part:BBa_K2347008">BBa_K2347008</a></td> | <td><a target="_blank" href="http://parts.igem.org/Part:BBa_K2347008">BBa_K2347008</a></td> | ||
<td>composite</td> | <td>composite</td> | ||
− | <td> | + | <td>NADH->NAD+</td> |
<td>2862bp</td> | <td>2862bp</td> | ||
</tr> | </tr> | ||
</tbody> | </tbody> | ||
</table> | </table> | ||
− | <h4>1.pSBC3-ADH1+gld+tGPD1<br><h4> | + | <h4> |
− | + | ||
+ | <div class="col-md-12" style="padding-top:30px"> | ||
+ | <div class="col-md-6"> | ||
+ | <center><h4>1.pSBC3-ADH1+gld+tGPD1<br><h4></center> | ||
+ | <center> <h4>In this part, the promoter of gld is ADH1 promoter, and the terminator is tGPD1 terminator. ADH1 promoter is | ||
a kind of promoter which has strong expression and tGPD1 terminator is a terminator with rather strong | a kind of promoter which has strong expression and tGPD1 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 | expression ability. So they can both increase the expression of genes. At the same time they are constitutive | ||
Line 227: | Line 231: | ||
yeast, thereby reducing the burden of plasmid expression in yeast. We linked this part with part | yeast, thereby reducing the burden of plasmid expression in yeast. We linked this part with part | ||
pSBC3-PGK1 + DAK + TPFK1 and the YCplac33 plasmid vector to form intact plasmid with URA deficient. | pSBC3-PGK1 + DAK + TPFK1 and the YCplac33 plasmid vector to form intact plasmid with URA deficient. | ||
− | </h4><br> | + | </h4><br></center> |
− | <h4>2.pSBC3-PGK1+DAK+ | + | <center> <img src="https://static.igem.org/mediawiki/2017/e/e5/NPU-pSB1C3-ADH1-GlyDH-tGPD1.png" class="img-responsive"></center> |
− | + | <h4> </h4> | |
− | is a kind of promoter which has strong expression and | + | </div> |
+ | |||
+ | |||
+ | <div class="col-md-6"> | ||
+ | |||
+ | <center> <h4>2.pSBC3-PGK1+DAK+TPFK1<br><h4></center> | ||
+ | <center> <h4>In this part, the promoter of DAK is PGK1 promoter, and the terminator is TPFK1 terminator. PGK1 promoter | ||
+ | is a kind of promoter which has strong expression and TPFK1 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 | 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, DAK, into the chromosome | promoters and terminators, which have the ability to integrate gene fragment, DAK, into the chromosome | ||
of yeast, thereby reducing the burden of plasmid expression in yeast. We linked this part with part | of yeast, thereby reducing the burden of plasmid expression in yeast. We linked this part with part | ||
pSBC3-ADH1+gld+tGPD1 and the YCplac33 plasmid vector to form intact plasmid with URA deficient. | pSBC3-ADH1+gld+tGPD1 and the YCplac33 plasmid vector to form intact plasmid with URA deficient. | ||
− | </h4><br> | + | </h4><br></center> |
− | <h4>3.pSBC3-pTDH3+ceas2+tPFK1<br><h4> | + | |
+ | <center> <img src="https://static.igem.org/mediawiki/2017/e/e8/PSB1C3-PGK1-DAK-tPFK1.png" class="img-responsive"></center> | ||
+ | <h4> </h4> | ||
+ | </div> | ||
+ | |||
+ | </div> | ||
+ | <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 | ||
+ | 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> | ||
+ | <center> <img src="https://static.igem.org/mediawiki/2017/6/65/YCplac33-URA-gld-DAK.png" class="img-responsive"></center> | ||
+ | |||
+ | </h4> | ||
+ | <h4>3.pSBC3-pTDH3+ceas2+tPFK1<br><h4> | ||
<h4>In this part, the promoter of ceaS2 is pTDH3 promoter, and the terminator is tPFK1 terminator. pTDH3 | <h4>In this part, the promoter of ceaS2 is pTDH3 promoter, and the terminator is tPFK1 terminator. pTDH3 | ||
promoter is a kind of promoter which has strong expression and tPFK1 terminator is a terminator with | promoter is a kind of promoter which has strong expression and tPFK1 terminator is a terminator with | ||
Line 244: | Line 271: | ||
with the YCplac33 plasmid vector to form intact plasmid with LEU deficient. | with 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"> | ||
+ | <div class="col-md-6"> | ||
+ | <img src="https://static.igem.org/mediawiki/2017/6/6c/PSB1C3-pTDH3%2Bceas2%2BtPFK1.png" class="img-responsive"> | ||
+ | <h4> </h4> | ||
+ | </div> | ||
+ | <div class="col-md-6"> | ||
+ | <img src="https://static.igem.org/mediawiki/2017/0/06/YCplac-pTDH3-ceaS2-tPFK1.png" class="img-responsive"> | ||
+ | <h4> </h4> | ||
+ | |||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | <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>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"> | ||
+ | <div class="col-md-6"> | ||
+ | <img src="https://static.igem.org/mediawiki/2017/c/c4/PSB1C3-TEF2-NOX-tRPS2.png" class="img-responsive"> | ||
+ | <h4> </h4> | ||
+ | </div> | ||
+ | <div class="col-md-6"> | ||
+ | <img src="https://static.igem.org/mediawiki/2017/8/86/YCplac33-Leu-NOX-ceaS2.png" class="img-responsive"> | ||
+ | <h4> </h4> | ||
− | + | </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> | ||
</div> | </div> | ||
Latest revision as of 02:33, 2 November 2017