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**Note: The word “Pantide” in the following paragraph refers to the collection of all the toxin design in our project. | **Note: The word “Pantide” in the following paragraph refers to the collection of all the toxin design in our project. | ||
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<p class="content-1">Testee selection</p> | <p class="content-1">Testee selection</p> | ||
<p class="content">In the test of Pantide toxicity, we chose tobacco cutworm as the testee for the insect appetite tests because all the Pantide targets lepidopterans. We hope that we can observe the difference of the three toxins in vivo. Tobacco cutworms, which are one of the major pests in vegetable farms, impact on Crucifers in farmland around the world. In the serious pest damage, the density of tobacco cutworms is approximately up to two hundred million per ha[1]. Therefore, we used five tobacco cutworms to mimic the much more severe situation in our insect experiment.</p> | <p class="content">In the test of Pantide toxicity, we chose tobacco cutworm as the testee for the insect appetite tests because all the Pantide targets lepidopterans. We hope that we can observe the difference of the three toxins in vivo. Tobacco cutworms, which are one of the major pests in vegetable farms, impact on Crucifers in farmland around the world. In the serious pest damage, the density of tobacco cutworms is approximately up to two hundred million per ha[1]. Therefore, we used five tobacco cutworms to mimic the much more severe situation in our insect experiment.</p> | ||
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<p class="content-1">Experiment Design</p> | <p class="content-1">Experiment Design</p> | ||
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In the following experiments, we used E. coli BL21 Rosetta-Gami strain to produce Pantide. We aimed to evaluate the remained leaf area applying Pantide. In this insect test, Pantide was diluted into three concentrations to observe the trends of dose response and to confirm the quality of PANTIDE. All the following experiments have three dilution ratios including 1/125 x, 1/25x and 1/5x. Three repeated tests were done in each experiment. See the procedure of insect experiment on Protocols. | In the following experiments, we used E. coli BL21 Rosetta-Gami strain to produce Pantide. We aimed to evaluate the remained leaf area applying Pantide. In this insect test, Pantide was diluted into three concentrations to observe the trends of dose response and to confirm the quality of PANTIDE. All the following experiments have three dilution ratios including 1/125 x, 1/25x and 1/5x. Three repeated tests were done in each experiment. See the procedure of insect experiment on Protocols. | ||
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<p class="content-1">Hypothesis</p> | <p class="content-1">Hypothesis</p> | ||
<p class="content">According to the mechanism of Pantide, Pantide is supposed to perform its toxicity in the nervous system of the larvae. We might see larvae paralyze and finally die. So that we may define Pantide as a biological pesticide.</p> | <p class="content">According to the mechanism of Pantide, Pantide is supposed to perform its toxicity in the nervous system of the larvae. We might see larvae paralyze and finally die. So that we may define Pantide as a biological pesticide.</p> | ||
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<p class="title">Insect Test Result</p> | <p class="title">Insect Test Result</p> | ||
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<div> | <div> | ||
<img src="https://static.igem.org/mediawiki/2016/e/e6/NCTU_FIG2.jpeg" class="picture"> | <img src="https://static.igem.org/mediawiki/2016/e/e6/NCTU_FIG2.jpeg" class="picture"> | ||
− | <p class="content-image" style="text-align:center !important;">Figure | + | <p class="content-image" style="text-align:center !important;">Figure 2. The analysis of remained leaf area in the pre-test with the Hv1a and Hv1a-lectin.</p> |
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+ | <p class="content">Figure 1. shows the pictures of the remained leaf disks after twelve hours of feeding assays. After we had done the feeding assays on tobacco cutworms with the three dilution ratio, we measured the area with the software imageJ. Figure 2. shows the percentage of the average remained area on the leaf disks. The higher the bar is, the larger the remained leaves area is. The observed phenomenon can be analyzed below.</p> | ||
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</section> | </section> |
Revision as of 12:22, 18 October 2016