Difference between revisions of "Team:NPU-China/CompositeParts"

 
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                     </tbody>
 
                     </tbody>
 
                     </table>
 
                     </table>
−
                     <h4>1.1pSBC3-ADH1+gld+tGPD1<br><h4>
+
<h4>
−
                    <h4>In this part, the promoter of gld is ADH1 promoter, and the terminator is tGPD1 terminator. ADH1 promoter is  
+
 
 +
              <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  
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                         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+TPFK1<br><h4>
+
                      <center>  <img src="https://static.igem.org/mediawiki/2017/e/e5/NPU-pSB1C3-ADH1-GlyDH-tGPD1.png" class="img-responsive"></center>
−
                    <h4>In this part, the promoter of DAK is PGK1 promoter, and the terminator is TPFK1 terminator. PGK1 promoter   
+
                        <h4> </h4>
 +
                     </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  
 
                         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   
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                         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  
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                         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  
+
            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
+
            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  
+
            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  
+
            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.
+
            pSBC3-pTDH3+ceas2+tPFK1and the YCplac33 plasmid vector to form intact plasmid with LEU deficient.
−
                        </h4><br>
+
−
                    <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
+
−
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>
+
−
                    <h3>How we use composite parts?<h3><br>
+
−
                    <h4>1.pSBC3-pTDH3+ceas2+tPFK1<br><h4>
+
−
                    <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>2.pSBC3-ADH1+gld+tGPD1 and pSBC3-PGK1+DAK+TPFK1<br><h4>
+
−
                    <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><br>
 
                         </h4><br>
−
                    <h4>3.pSBC3-TEF2+NOX+tRPS2<br><h4>
+
        <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
+
            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.
+
            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

Name Type Description Length
BBa_K2347005 composite DHAP/G3P->acrylic acid 2862bp
BBa_K2347006 composite glycerol->DHA 2862bp
BBa_K2347007 composite DHA->DHAP 2862bp
BBa_K2347008 composite NADH->NAD+ 2862bp

1.pSBC3-ADH1+gld+tGPD1

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 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, gld, into the chromosome of 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.


2.pSBC3-PGK1+DAK+TPFK1

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 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 pSBC3-ADH1+gld+tGPD1 and the YCplac33 plasmid vector to form intact plasmid with URA deficient.


pSBC3-ADH1+gld+tGPD1 & pSBC3-PGK1+DAK+TPFK1

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.

3.pSBC3-pTDH3+ceas2+tPFK1

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 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, ceaS2, into the chromosome of yeast, thereby reducing the burden of plasmid expression in yeast. We linked this part with the YCplac33 plasmid vector to form intact plasmid with LEU deficient.


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.


4.pSBC3-TEF2+NOX+tRPS2

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.


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.