Difference between revisions of "Team:CCU Taiwan/Composite Part"

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   This part contain a Kozak sequence for initiating and enhancing translation, Prepro-alpha-factor as a signal peptide for secretion, codon-optimized version of laccase gene sequence, HA tag for Western Blot protein detection and 6xHis-tag for protein purification.  
 
   This part contain a Kozak sequence for initiating and enhancing translation, Prepro-alpha-factor as a signal peptide for secretion, codon-optimized version of laccase gene sequence, HA tag for Western Blot protein detection and 6xHis-tag for protein purification.  
 
</p>
 
</p>
 +
 +
        <div id="Composite1" class="polaroid">
 +
                  <img src="https://static.igem.org/mediawiki/parts/5/59/T--CCU_Taiwan--111.png" style="width:100%">
 +
                  <div class="container">
 +
                    <p>Figure 1: The schematic structure of gene design- Lac1.</p>
 +
                  </div>
 +
                </div>
 +
              <p class="description"><strong>Part construction in pGAPZ A:</strong><br>
 +
 +
        <div id="Composite2" class="polaroid">
 +
                  <img src="https://static.igem.org/mediawiki/parts/f/f2/T--CCU_Taiwan--_pGAPZ_A_Lac1%2C_pGAPZ_A_Px16%2C_pGAPZ_A_Px18_Colony_PCR.jpeg" style="width:100%">
 +
                  <div class="container">
 +
                    <p>Figure 2: PCR result. <br>Well1: Marker/ Well2, 3, 4: Lac1 Colony 1, 2, 3/ Well5, 6, 7: Px16 Colony 1, 2, 3/ Well8, 9, 10: Px18 Colony 1, 2, 3/ Well11: pGAPZ A Colony
 +
<br>
 +
Insert long after PCR — pGAPZ A_Px16 :1377 bp/ pGAPZ A_Px18 :1359 bp/ pGAPZ A_Lac1 :2124bp
 +
</p>
 +
                  </div>
 +
                </div>
 +
        <div id="Composite3" class="polaroid">
 +
                  <img src="https://static.igem.org/mediawiki/parts/a/ae/T--CCU_Taiwan--_pSB1C3_Lac1%2C_pSB1C3_Px16%2C_pSB1C3_Px18_Miniprep_and_digest_with_EcoRI%2CAgeI.jpeg" style="width:100%">
 +
                  <div class="container">
 +
                    <p>Figure 3: Miniprep plasmid digest by EcoRI, PstI.<br> WWell 1: Marker/ Well2, 3, 4: Plasmid of Lac1 colony1, 2, 3 after EcoRI, AgeI digest/ Well5, 6, 7: Plasmid of Px16 colony1, 2, 3 after EcoRI, AgeI digest/ Well8, 9, 10: Plasmid of Px18 colony1, 2, 3 after EcoRI, AgeI digest
 +
<br>
 +
Vector long after digest — pGAPZ A: 2884bp
 +
<br>
 +
Insert long after digest — Px16: 1295bp/ Px18: 1277bp/ Lac1: 2042bp
 +
</p>
 +
                  </div>
 +
                </div>
 +
 +
 +
 
<p class="second">BBa_K2809021<br>
 
<p class="second">BBa_K2809021<br>
 
peroxidase 18
 
peroxidase 18
Line 165: Line 197:
 
   
 
   
 
</p>
 
</p>
 +
        <div id="Composite4" class="polaroid">
 +
                  <img src="https://static.igem.org/mediawiki/parts/9/9e/T--CCU_Taiwan--114.png" style="width:100%">
 +
                  <div class="container">
 +
                    <p>Figure 4: The schematic structure of gene design- Px18.</p>
 +
                  </div>
 +
                </div><br><br>
 +
 +
        <div id="Composite5" class="polaroid">
 +
                  <img src="https://static.igem.org/mediawiki/parts/f/f2/T--CCU_Taiwan--_pGAPZ_A_Lac1%2C_pGAPZ_A_Px16%2C_pGAPZ_A_Px18_Colony_PCR.jpeg" style="width:100%">
 +
                  <div class="container">
 +
                    <p>Figure 5: PCR result.<br> Well1: Marker/ Well2, 3, 4: Lac1 Colony 1, 2, 3/ Well5, 6, 7: Px16 Colony 1, 2, 3/ Well8, 9, 10: Px18 Colony 1, 2, 3/ Well11: pGAPZ A Colony
 +
<br>
 +
Insert long after PCR — pGAPZ A_Px16 :1377 bp/ pGAPZ A_Px18 :1359 bp/ pGAPZ A_Lac1 :2124bp
 +
</p>
 +
                  </div>
 +
                </div>
 +
        <div id="Composite6" class="polaroid">
 +
                  <img src="https://static.igem.org/mediawiki/parts/a/ae/T--CCU_Taiwan--_pSB1C3_Lac1%2C_pSB1C3_Px16%2C_pSB1C3_Px18_Miniprep_and_digest_with_EcoRI%2CAgeI.jpeg" style="width:100%">
 +
                  <div class="container">
 +
                    <p>Figure 6: Miniprep plasmid digest by EcoRI, PstI.<br>Well 1: Marker/ Well2, 3, 4: Plasmid of Lac1 colony1, 2, 3 after EcoRI, AgeI digest/ Well5, 6, 7: Plasmid of Px16 colony1, 2, 3 after EcoRI, AgeI digest/ Well8, 9, 10: Plasmid of Px18 colony1, 2, 3 after EcoRI, AgeI digest
 +
<br>
 +
Vector long after digest — pGAPZ A: 2884bp
 +
<br>
 +
Insert long after digest — Px16: 1295bp/ Px18: 1277bp/ Lac1: 2042bp
 +
</p>
 +
                  </div>
 +
                </div><br>
 
<p class="second">BBa_K2809031<br>
 
<p class="second">BBa_K2809031<br>
 
peroxidase 16
 
peroxidase 16
Line 171: Line 230:
 
   This part contain a Kozak sequence for initiating and enhancing translation, Prepro-alpha-factor as a signal peptide for secretion, codon-optimized version of peroxidase 16 gene sequence, FLAG-tag for Western Blot protein detection and 6xHis-tag for protein purification.  
 
   This part contain a Kozak sequence for initiating and enhancing translation, Prepro-alpha-factor as a signal peptide for secretion, codon-optimized version of peroxidase 16 gene sequence, FLAG-tag for Western Blot protein detection and 6xHis-tag for protein purification.  
  
−
</p><br><br><br>
+
</p><br>         <div id="Composite7" class="polaroid">
 +
                  <img src="https://static.igem.org/mediawiki/parts/1/1e/T--CCU_Taiwan--117.png" style="width:100%">
 +
                  <div class="container">
 +
                    <p>Figure 7: The schematic structure of gene design- Px16.</p>
 +
                  </div>
 +
                </div><br>
 +
 
 +
        <div id="Composite8" class="polaroid">
 +
                  <img src="https://static.igem.org/mediawiki/parts/f/f2/T--CCU_Taiwan--_pGAPZ_A_Lac1%2C_pGAPZ_A_Px16%2C_pGAPZ_A_Px18_Colony_PCR.jpeg" style="width:100%">
 +
                  <div class="container">
 +
                    <p>Figure 8: PCR result.<br> Well1: Marker/ Well2, 3, 4: Lac1 Colony 1, 2, 3/ Well5, 6, 7: Px16 Colony 1, 2, 3/ Well8, 9, 10: Px18 Colony 1, 2, 3/ Well11: pGAPZ A Colony
 +
<br>
 +
Insert long after PCR — pGAPZ A_Px16 :1377 bp/ pGAPZ A_Px18 :1359 bp/ pGAPZ A_Lac1 :2124bp
 +
</p>
 +
                  </div>
 +
                </div>
 +
        <div id="Composite9" class="polaroid">
 +
                  <img src="https://static.igem.org/mediawiki/parts/a/ae/T--CCU_Taiwan--_pSB1C3_Lac1%2C_pSB1C3_Px16%2C_pSB1C3_Px18_Miniprep_and_digest_with_EcoRI%2CAgeI.jpeg" style="width:100%">
 +
                  <div class="container">
 +
                    <p>Figure 9: Miniprep plasmid digest by EcoRI, PstI.<br>Well 1: Marker/ Well2, 3, 4: Plasmid of Lac1 colony1, 2, 3 after EcoRI, AgeI digest/ Well5, 6, 7: Plasmid of Px16 colony1, 2, 3 after EcoRI, AgeI digest/ Well8, 9, 10: Plasmid of Px18 colony1, 2, 3 after EcoRI, AgeI digest
 +
<br>
 +
Vector long after digest — pGAPZ A: 2884bp
 +
<br>
 +
Insert long after digest — Px16: 1295bp/ Px18: 1277bp/ Lac1: 2042bp
 +
</p>
 +
                  </div>
 +
                </div>
 +
 
 +
<br>
 +
<br>
 +
<br>
 +
 
 
</div>
 
</div>
 
</div>
 
</div>

Revision as of 18:13, 17 October 2018

COMPOSITE PART

BBa_K2809011
laccase 1

We use pGAPZ A vector from invitrogen and insert this part to construct our vector. We constructed this part by combining BBa_K2809001, BBa_K2809002 and BBa_K2809010.
This part contain a Kozak sequence for initiating and enhancing translation, Prepro-alpha-factor as a signal peptide for secretion, codon-optimized version of laccase gene sequence, HA tag for Western Blot protein detection and 6xHis-tag for protein purification.

Figure 1: The schematic structure of gene design- Lac1.

Part construction in pGAPZ A:

Figure 2: PCR result.
Well1: Marker/ Well2, 3, 4: Lac1 Colony 1, 2, 3/ Well5, 6, 7: Px16 Colony 1, 2, 3/ Well8, 9, 10: Px18 Colony 1, 2, 3/ Well11: pGAPZ A Colony
Insert long after PCR — pGAPZ A_Px16 :1377 bp/ pGAPZ A_Px18 :1359 bp/ pGAPZ A_Lac1 :2124bp

Figure 3: Miniprep plasmid digest by EcoRI, PstI.
WWell 1: Marker/ Well2, 3, 4: Plasmid of Lac1 colony1, 2, 3 after EcoRI, AgeI digest/ Well5, 6, 7: Plasmid of Px16 colony1, 2, 3 after EcoRI, AgeI digest/ Well8, 9, 10: Plasmid of Px18 colony1, 2, 3 after EcoRI, AgeI digest
Vector long after digest — pGAPZ A: 2884bp
Insert long after digest — Px16: 1295bp/ Px18: 1277bp/ Lac1: 2042bp

BBa_K2809021
peroxidase 18

We use pGAPZ A vector from invitrogen and insert this part to construct our vector. We constructed this part by combining BBa_K2809001, BBa_K2809002 and BBa_K2809020.
This part contain a Kozak sequence for initiating and enhancing translation, Prepro-alpha-factor as a signal peptide for secretion, codon-optimized version of peroxidase 18 gene sequence, V5-tag for Western Blot protein detection and 6xHis-tag for protein purification.

Figure 4: The schematic structure of gene design- Px18.



Figure 5: PCR result.
Well1: Marker/ Well2, 3, 4: Lac1 Colony 1, 2, 3/ Well5, 6, 7: Px16 Colony 1, 2, 3/ Well8, 9, 10: Px18 Colony 1, 2, 3/ Well11: pGAPZ A Colony
Insert long after PCR — pGAPZ A_Px16 :1377 bp/ pGAPZ A_Px18 :1359 bp/ pGAPZ A_Lac1 :2124bp

Figure 6: Miniprep plasmid digest by EcoRI, PstI.
Well 1: Marker/ Well2, 3, 4: Plasmid of Lac1 colony1, 2, 3 after EcoRI, AgeI digest/ Well5, 6, 7: Plasmid of Px16 colony1, 2, 3 after EcoRI, AgeI digest/ Well8, 9, 10: Plasmid of Px18 colony1, 2, 3 after EcoRI, AgeI digest
Vector long after digest — pGAPZ A: 2884bp
Insert long after digest — Px16: 1295bp/ Px18: 1277bp/ Lac1: 2042bp


BBa_K2809031
peroxidase 16

We use pGAPZ A vector from invitrogen and insert this part to construct our vector. We constructed this part by combining BBa_K2809001, BBa_K2809002 and BBa_K2809030.
This part contain a Kozak sequence for initiating and enhancing translation, Prepro-alpha-factor as a signal peptide for secretion, codon-optimized version of peroxidase 16 gene sequence, FLAG-tag for Western Blot protein detection and 6xHis-tag for protein purification.


Figure 7: The schematic structure of gene design- Px16.


Figure 8: PCR result.
Well1: Marker/ Well2, 3, 4: Lac1 Colony 1, 2, 3/ Well5, 6, 7: Px16 Colony 1, 2, 3/ Well8, 9, 10: Px18 Colony 1, 2, 3/ Well11: pGAPZ A Colony
Insert long after PCR — pGAPZ A_Px16 :1377 bp/ pGAPZ A_Px18 :1359 bp/ pGAPZ A_Lac1 :2124bp

Figure 9: Miniprep plasmid digest by EcoRI, PstI.
Well 1: Marker/ Well2, 3, 4: Plasmid of Lac1 colony1, 2, 3 after EcoRI, AgeI digest/ Well5, 6, 7: Plasmid of Px16 colony1, 2, 3 after EcoRI, AgeI digest/ Well8, 9, 10: Plasmid of Px18 colony1, 2, 3 after EcoRI, AgeI digest
Vector long after digest — pGAPZ A: 2884bp
Insert long after digest — Px16: 1295bp/ Px18: 1277bp/ Lac1: 2042bp