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<div id="pt"> | <div id="pt"> | ||
− | <h8>PRK</h8></br></br> | + | <h8>PRK (PHOSPHORIBULOKINASE)</h8></br></br> |
<p class="pcontent"> | <p class="pcontent"> | ||
Achievements:</br> | Achievements:</br> | ||
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<img class="contentimg fig5" src="https://static.igem.org/mediawiki/2018/f/f6/T--NCKU_Tainan--Results_fig_4_2.PNG"> | <img class="contentimg fig5" src="https://static.igem.org/mediawiki/2018/f/f6/T--NCKU_Tainan--Results_fig_4_2.PNG"> | ||
− | <p class=" | + | <p class="pcenter"> |
Fig 5. The result of PRK test in W3110 | Fig 5. The result of PRK test in W3110 | ||
</p></br> | </p></br> | ||
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<div id="pt"> | <div id="pt"> | ||
− | <h8>CA</h8></br></br> | + | <h8>CA (Carbonic Anhydrase)</h8></br></br> |
<p class="pcontent"> | <p class="pcontent"> | ||
Achievements:</br> | Achievements:</br> | ||
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<div id="pt"> | <div id="pt"> | ||
<p class="pcontent"> | <p class="pcontent"> | ||
− | The engineered <i>E. coli</i> BL21 (DE3) | + | The engineered <i>E. coli</i> BL21 (DE3) is cultured in M9 medium with formula |
adjusted so that xylose is the sole carbon source. The aforementioned M9 Medium | adjusted so that xylose is the sole carbon source. The aforementioned M9 Medium | ||
contains | contains | ||
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$${=\ 0.575\ {mg\over L ∙hr}}$$ | $${=\ 0.575\ {mg\over L ∙hr}}$$ | ||
− | To find out how much carbon in biomass comes from the carbon in | + | To find out how much carbon in biomass comes from the carbon in CO2 captured by the heterotrophic microbes, we can divide equation (3) by the mass percentage of carbon in biomass: |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | </p> | |
− | + | <p class="pcontent">$${{{ \{ CO_{2 net}} \} \over \{ {C_{biomass}} \} } \geq {1 - | |
+ | { \{ {C_{xylose}} \} \over \{ {C_{biomass}} \} }}}$$</p> | ||
+ | <p class="pcontent">We can thus calculate the ratio with our experiment results:</p> | ||
+ | <p class="pcontent">$${{Ratio \ of \ carbon \ in \ CO_2 \ fixed \ to \ carbon \ in \ biomass} = {1 -{0.0689 \over 0.0758}} = 9.1 \%}$$ | ||
</p> | </p> | ||
</div> | </div> | ||
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modified with 1M HCl). We then took the sample and incubate in the 96 well and | modified with 1M HCl). We then took the sample and incubate in the 96 well and | ||
measure its fluorescence intensity for every 3 minutes. We found out that the | measure its fluorescence intensity for every 3 minutes. We found out that the | ||
− | promoter | + | promoter P<sub>asr</sub> will be induced at the pH value below four within 30 minutes. The |
different fluorescence intensity can be observed within 30 minutes. The | different fluorescence intensity can be observed within 30 minutes. The | ||
fluorescence had the peak at pH value of 4.25. | fluorescence had the peak at pH value of 4.25. | ||
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<p class="pcontent"> | <p class="pcontent"> | ||
− | Based on the data has shown above, we could conclude that | + | Based on the data has shown above, we could conclude that P<sub>asr</sub> is an acidic |
promoter as it has a high expression of fluorescence at pH 4.25 and pH 5. The | promoter as it has a high expression of fluorescence at pH 4.25 and pH 5. The | ||
− | results show that | + | results show that P<sub>asr</sub> constructed pH sensing system can be used as an alert. |
When the medium turns acidic, fluorescence can be easily observed. We believe | When the medium turns acidic, fluorescence can be easily observed. We believe | ||
that this system can also be applied to various bio-detection system. | that this system can also be applied to various bio-detection system. | ||
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modified with 1M HCl). The induction of P<sub>gadA</sub> is observed under | modified with 1M HCl). The induction of P<sub>gadA</sub> is observed under | ||
neutral and | neutral and | ||
− | + | weak acidic environment. | |
</p> | </p> | ||
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P<sub>asr</sub> and would like to improve the sensitivity of this biobrick. We | P<sub>asr</sub> and would like to improve the sensitivity of this biobrick. We | ||
thus add a | thus add a | ||
− | RiboJ sequence at the downstream of P<sub>gadA</sub>. RiboJ sequence is reported to increase the expression of downstream protein. We thus compare the | + | RiboJ sequence at the downstream of P<sub>gadA</sub>. RiboJ sequence is reported to increase the expression of downstream protein. We thus compare the fluorescence of previous and improved biobrick. We discover that with RiboJ, the protein of down-stream reporter protein is increased.</p> |
<img class="contentimg fig3" src="https://static.igem.org/mediawiki/2018/f/fd/T--NCKU_Tainan--PGADA_OMPARISON.png"> | <img class="contentimg fig3" src="https://static.igem.org/mediawiki/2018/f/fd/T--NCKU_Tainan--PGADA_OMPARISON.png"> |
Latest revision as of 03:26, 18 October 2018
Results
Hard Work Pays Off