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Team:Hannover/Results/Heavy Metals/Arabidopsis
From 2014.igem.org
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<h2>Results</h2> | <h2>Results</h2> | ||
| - | <p class="text">The exchange of the promoter was checked by sequencing. We achieved to regenerate young <i>A. thaliana</i> after the transformation with our T4MBP. To receive a stabile F2 generation seeds of transformed plants would have to be harvested and grown. As a next step of this test series the plants would have been transfered on medium with heavy metals. In a following analysis it would be detected if these plants bind the heavy metal zinc, copper, cadmium and/or arsenic. </p> | + | <p class="text">The exchange of the promoter was checked by sequencing. We achieved to regenerate young <i>A. thaliana</i> after the transformation with our T4MBP. A PCR and a immunoblot would indicate if tested plant were positive or negative for our T4MBP. To receive a stabile F2 generation seeds of transformed plants would have to be harvested and grown. As a next step of this test series the plants would have been transfered on medium with heavy metals. In a following analysis it would be detected if these plants bind the heavy metal zinc, copper, cadmium and/or arsenic. </p> |
<center><table border="0"><tr><td><a href="https://static.igem.org/mediawiki/2014/6/6a/Hannover_20141012_Arabudopsis-results1.jpg" data-lightbox="galery1" data-title="Fig. 1a: Young potentially transgenic <i>A. thaliana</i> during the transfer from medium to earth. "><img src="https://static.igem.org/mediawiki/2014/6/6a/Hannover_20141012_Arabudopsis-results1.jpg" width="300px"></a></td><td><a href="https://static.igem.org/mediawiki/2014/1/13/Hannover_20141012_Arabudopsis-results3.jpg | <center><table border="0"><tr><td><a href="https://static.igem.org/mediawiki/2014/6/6a/Hannover_20141012_Arabudopsis-results1.jpg" data-lightbox="galery1" data-title="Fig. 1a: Young potentially transgenic <i>A. thaliana</i> during the transfer from medium to earth. "><img src="https://static.igem.org/mediawiki/2014/6/6a/Hannover_20141012_Arabudopsis-results1.jpg" width="300px"></a></td><td><a href="https://static.igem.org/mediawiki/2014/1/13/Hannover_20141012_Arabudopsis-results3.jpg | ||
" data-lightbox="galery1" data-title="Fig. 1b: Young potentially transgenic <i>A. thaliana</i> in medium."><img src="https://static.igem.org/mediawiki/2014/1/13/Hannover_20141012_Arabudopsis-results3.jpg" width="330px"></a></td><td><a href="https://static.igem.org/mediawiki/2014/3/35/Hannover_20141012_Arabudopsis-results2.jpg | " data-lightbox="galery1" data-title="Fig. 1b: Young potentially transgenic <i>A. thaliana</i> in medium."><img src="https://static.igem.org/mediawiki/2014/1/13/Hannover_20141012_Arabudopsis-results3.jpg" width="330px"></a></td><td><a href="https://static.igem.org/mediawiki/2014/3/35/Hannover_20141012_Arabudopsis-results2.jpg | ||
Revision as of 22:24, 16 October 2014
Results / Heavy metals / Arabidopsis thaliana T4MBP
Labwork
Transformation of A. thaliana with our T4MBP:
- Exchange the original promoter enTCUP2 of the binary vector pORE_E3 (AY562536.1) into a 2x35s promoter using resctriction site based cloning.
- We integrated our T4MBP in our modified pORE_E3_2x35S.
- Application of our floral dip method.
- After 6 weeks we harvested transgenic seeds from A. thaliana and plated these seeds on selection MSO-media.
- At last we potted transformed plants.
Results
The exchange of the promoter was checked by sequencing. We achieved to regenerate young A. thaliana after the transformation with our T4MBP. A PCR and a immunoblot would indicate if tested plant were positive or negative for our T4MBP. To receive a stabile F2 generation seeds of transformed plants would have to be harvested and grown. As a next step of this test series the plants would have been transfered on medium with heavy metals. In a following analysis it would be detected if these plants bind the heavy metal zinc, copper, cadmium and/or arsenic.
 |  |  |
Fig. 1: Pictures showing young potentially transgenic A. thaliana before, during and after the transfer from medium to earth. | ||
|---|---|---|
Below in figures 2 and 3 you can see the original vector pORE_E3 with an enTCUP2 promoter. For a better expression of our T4MBP protein we exchanged the enTCUP2 with the 2x35S promoter. Each step of this procedure is visualized in this histroy . Furthermore this history includes the insertion of our T4MBP (there termed CDS for coding sequence).
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Fig. 2: Vector pORE_E3 with the original enTCUP2 promoter. | Fig. 3: Vector pORE-E3 with 2x35S promoter and our T4MBP which includes a sequence for expansin 4, cellulose-binding domain and domains for the binding of copper, arsenic, zinc, cadmium. |
