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Team:USTC CHINA/Project/Results
From 2013.igem.org
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<h1>Results</h1> | <h1>Results</h1> | ||
| + | <h2>Verifying the validity of our circuit by GFP</h2> | ||
<p>① E.coli BL21 proved that the standard transdermal locus does work with GFP.</p> | <p>① E.coli BL21 proved that the standard transdermal locus does work with GFP.</p> | ||
<img src="https://static.igem.org/mediawiki/2013/8/82/2013ustc-china_genecircuit5.png" width="580" height="160"/> | <img src="https://static.igem.org/mediawiki/2013/8/82/2013ustc-china_genecircuit5.png" width="580" height="160"/> | ||
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| - | <div class="bassic-bar"><p>② The expression of recombinant antigen and adjuvant in E.coli BL21.</p> | + | <div class="bassic-bar"> |
| + | <h2>TD1-fusion protein expression</h2> | ||
| + | <p>② The expression of recombinant antigen and adjuvant in E.coli BL21.</p> | ||
<img src="https://static.igem.org/mediawiki/2013/4/4d/2013ustc-china_genecircuit.png" width="580" height="100"/> | <img src="https://static.igem.org/mediawiki/2013/4/4d/2013ustc-china_genecircuit.png" width="580" height="100"/> | ||
<p>The practicality of our locus afforded by TD1-GFP enabled our to express recombinant antigen TD1-HBsAg and recombinant adjuvant TD1-LTB successfully. So far our basic molecule experiments have ended with perfection.</p> | <p>The practicality of our locus afforded by TD1-GFP enabled our to express recombinant antigen TD1-HBsAg and recombinant adjuvant TD1-LTB successfully. So far our basic molecule experiments have ended with perfection.</p> | ||
<div style="float:left;width:200px;"> | <div style="float:left;width:200px;"> | ||
| - | <img src="https://static.igem.org/mediawiki/2013/6/6f/2013ustc-chinasdsTD1-LTB.png" width="200" height=" | + | <img src="https://static.igem.org/mediawiki/2013/6/6f/2013ustc-chinasdsTD1-LTB.png" width="200" height="300"/> |
</div> | </div> | ||
<div style="float:left;width:380px;"> | <div style="float:left;width:380px;"> | ||
| - | <img src="https://static.igem.org/mediawiki/2013/7/73/2013ustc-chinasdsTD1-HBsAg.png" width="380" height=" | + | <img src="https://static.igem.org/mediawiki/2013/7/73/2013ustc-chinasdsTD1-HBsAg.png" width="380" height="300"/> |
</div> | </div> | ||
<div class="atfigure" align="center">Fig3. SDS PAGE shows the expression of LTB(L)、HBsAg(R)</div> | <div class="atfigure" align="center">Fig3. SDS PAGE shows the expression of LTB(L)、HBsAg(R)</div> | ||
</div> | </div> | ||
| - | <div class="bassic-bar"><p>③ Verified the antigenicity of TD1-HBsAg by ELISA. The antigenicity of the TD1-antigen(fusion protein) strongly proved our theoretical basis. </p> | + | <div class="bassic-bar"> |
| - | <img src=" | + | <h2>Verified the antigenicity of TD1-antibody by ELISA</h2> |
| - | + | <p>③ Verified the antigenicity of TD1-HBsAg by ELISA. The antigenicity of the TD1-antigen(fusion protein)strongly proved our theoretical basis. </p> | |
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Revision as of 00:32, 27 September 2013
Stage one：basal experiments
Introduction
As the construction and concentration of recombinant plasmid rely heavily on E.coli system in current molecule experiment protocols of Bacillus subtilis, Bacillus subtilis acts only as the secretory expression vector. Therefore, to verify the practicality of our locus and the transdermal function of recombinant transdermal protein on top of its original functions, we conducted basic experiments on E.coli to verify our assumptions.
The following figure shows the stages of our basal experiments:
Results
Verifying the validity of our circuit by GFP
① E.coli BL21 proved that the standard transdermal locus does work with GFP.
Using GFP to prove the validity of a newly designed circuit is a classical way to verify the expressing of this circuit. As expressions in E.coli involve neither secretory nor sequential problems, we hoped to verify the practicality of our locus by the expression of TD1-GFP. Thus we selected pET22b,which is a common recombinant vector for plasmid construction, as our recombinant vector and E.coli BL21 as engineered bacteria . We fused sequence TD1-GFP with T7 promoter from pET22b downstream and succeeded in expressing fusion protein TD1-GFP, induced by IPTG.
Fig2. The expression of GFP under fluorescence microscope
TD1-fusion protein expression
② The expression of recombinant antigen and adjuvant in E.coli BL21.
The practicality of our locus afforded by TD1-GFP enabled our to express recombinant antigen TD1-HBsAg and recombinant adjuvant TD1-LTB successfully. So far our basic molecule experiments have ended with perfection.
Verified the antigenicity of TD1-antibody by ELISA
③ Verified the antigenicity of TD1-HBsAg by ELISA. The antigenicity of the TD1-antigen(fusion protein)strongly proved our theoretical basis.
