Difference between revisions of "Team:ECUST/Biocide"

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<p>We insert fragment of Cecropin AD into vector pET-28a(Figure 1) </p>
 
<p>We insert fragment of Cecropin AD into vector pET-28a(Figure 1) </p>
 
<figure>
 
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       <img src="https://static.igem.org/mediawiki/2018/a/a5/T--ECUST--Biocide1.jpg" class="zoom">
 
       <img src="https://static.igem.org/mediawiki/2018/a/a5/T--ECUST--Biocide1.jpg" class="zoom">
 
       <figcaption><b>Figure 1. The vector pET-28a Vector is cut by NcoI and BamHI. Sequence of AD was chemically synthesized and amplified by PCR, then ligated with linearized vector by Ezmax.</b></figcaption>         
 
       <figcaption><b>Figure 1. The vector pET-28a Vector is cut by NcoI and BamHI. Sequence of AD was chemically synthesized and amplified by PCR, then ligated with linearized vector by Ezmax.</b></figcaption>         
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<p>The plasmid was transformed to E. coli BL21 and cultured at 37 °C for 12 h. positive monoclonal bacteria were cultured and verified by PCR(Figure 2). </p>
 
<p>The plasmid was transformed to E. coli BL21 and cultured at 37 °C for 12 h. positive monoclonal bacteria were cultured and verified by PCR(Figure 2). </p>
 
<figure>
 
<figure>
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<figure class="makeresponsive floatleft" style="width: 50%;">
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<figure class="makeresponsive floatleft" style="width: 50%;margin-left:30%;">
 
       <img src="https://static.igem.org/mediawiki/2018/c/c6/T--ECUST--Biocide2.jpg" class="zoom">
 
       <img src="https://static.igem.org/mediawiki/2018/c/c6/T--ECUST--Biocide2.jpg" class="zoom">
 
       <figcaption><b>Figure 2. 1% Agarose Gel Electrophoresis of PCR, which shows that our vector was successfully constructed.</b></figcaption>     
 
       <figcaption><b>Figure 2. 1% Agarose Gel Electrophoresis of PCR, which shows that our vector was successfully constructed.</b></figcaption>     
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<p>We verified the expression of Cecropin AD by SDS-PAGE(Figure 3). </p>
 
<p>We verified the expression of Cecropin AD by SDS-PAGE(Figure 3). </p>
 
<figure>
 
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<figure class="makeresponsive floatleft" style="width: 50%;">
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<figure class="makeresponsive floatleft" style="width: 50%;margin-left:30%;">
 
       <img src="https://static.igem.org/mediawiki/2018/8/86/T--ECUST--Biocide3.jpg" class="zoom">
 
       <img src="https://static.igem.org/mediawiki/2018/8/86/T--ECUST--Biocide3.jpg" class="zoom">
 
       <figcaption><b>Figure 3. The SDS-PAGE of Cecropin AD
 
       <figcaption><b>Figure 3. The SDS-PAGE of Cecropin AD
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<p>To test the effect of cecropin AD, we measured the growth curve of iron bacteria adding with different concentration of cecropin AD.</p>
 
<p>To test the effect of cecropin AD, we measured the growth curve of iron bacteria adding with different concentration of cecropin AD.</p>
 
<figure>
 
<figure>
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<figure class="makeresponsive floatleft" style="width: 50%;">
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<figure class="makeresponsive floatleft" style="width: 50%;margin-left:32%;">
 
       <img src="https://static.igem.org/mediawiki/2018/3/30/T--ECUST--Biocide4.jpg" class="zoom">
 
       <img src="https://static.igem.org/mediawiki/2018/3/30/T--ECUST--Biocide4.jpg" class="zoom">
 
       <figcaption><b>Figure 4. Growth curve of iron bacteria adding with different concentration of cecropin AD</b></figcaption>         
 
       <figcaption><b>Figure 4. Growth curve of iron bacteria adding with different concentration of cecropin AD</b></figcaption>         

Revision as of 03:19, 18 October 2018

BIOCIDE

Description

We insert fragment of Cecropin AD into vector pET-28a(Figure 1)

Figure 1. The vector pET-28a Vector is cut by NcoI and BamHI. Sequence of AD was chemically synthesized and amplified by PCR, then ligated with linearized vector by Ezmax.

The plasmid was transformed to E. coli BL21 and cultured at 37 °C for 12 h. positive monoclonal bacteria were cultured and verified by PCR(Figure 2).

Figure 2. 1% Agarose Gel Electrophoresis of PCR, which shows that our vector was successfully constructed.

We verified the expression of Cecropin AD by SDS-PAGE(Figure 3).

Figure 3. The SDS-PAGE of Cecropin AD Lane 1 :before induction Lane 2,3:after induction

To test the effect of cecropin AD, we measured the growth curve of iron bacteria adding with different concentration of cecropin AD.

Figure 4. Growth curve of iron bacteria adding with different concentration of cecropin AD

The figure shows that cecropin AD has powerful bacterividal activity. Only concentration of cecropin AD is low enough, can iron bacteria grow.