Team:UESTC-China/Project

From 2014.igem.org

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<p style="color:#1b1b1b;">In total, we constructed 11 different vectors, including two backbones, six mono-gene expression vectors and three multi-gene expression vectors. The production of HPS, PHI, and FDH are located in chloroplast, while FALDH is located in cytoplasm. Chloroplast transit peptides were used for the purpose of chloroplast orientation. For comparison, those vectors carrying the genes of HPS, PHI, and FDH without the presence of transit peptide were also constructed. Those genes are inserted into tobacco via Agrobacterium-mediated leaf disk transformation method. By performing DNA and RT-PCR analysis, we got about 30 positive plants for each vector. The formaldehyde absorbance ability of our super plants was explored by both qualitative and quantitative detection. The results showed that our super plants have remarkable increased abilities of formaldehyde tolerance and can dramatically reduce the concentration of air formaldehyde (Fig. 4). Due to the time limitation, the following investigations are under the way: 1) the effect of individual four key enzymes on the metabolic efficiency of formaldehyde; 2) whether the presence of transit peptides can affect the metabolic efficiency of formaldehyde; 3) the expression of the gene AHA2 in tobacco.</p><br/>
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<p style="color:#1b1b1b;">In total, we constructed 11 different vectors, including two backbones, six mono-gene expression vectors and three multi-gene expression vectors. The production of HPS, PHI, and FDH are located in chloroplast, while FALDH is located in cytoplasm. Chloroplast transit peptides were used for the purpose of chloroplast orientation. For comparison, those vectors carrying the genes of HPS, PHI, and FDH without the presence of transit peptide were also constructed. Those genes are inserted into tobacco via Agrobacterium-mediated leaf disk transformation method. By performing DNA and RT-PCR analysis, we got about 30 positive plants for each vector. The formaldehyde absorbance ability of our super plants was explored by both qualitative and quantitative detection. The results showed that our super plants have remarkable increased abilities of formaldehyde tolerance and can dramatically reduce the concentration of air formaldehyde (Fig. 4). Due to the time limitation, the following investigations are under the way: 1) the effect of individual four key enzymes on the metabolic efficiency of formaldehyde; 2) whether the presence of transit peptides can affect the metabolic efficiency of formaldehyde; 3) the expression of the gene AHA2 in tobacco.</p><br/>
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<strong>Fig.4</strong> Fig. 4 Phenotype testing of transgenic seedlings and wild-type on formaldehyde exposure. A: Before exposure to HCHO. B: Exposure to 20 μl 37% HCHO for one week.
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<strong>Fig.4</strong> Phenotype testing of transgenic and wildtype plants on formaldehyde exposure. A: Before exposure to HCHO. B: Exposure to 10 μl 37% HCHO for one week.
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Revision as of 14:57, 17 October 2014

UESTC-China