Difference between revisions of "Team:NCKU Tainan/Results"

 
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                                 <div id="pt">
 
                                 <div id="pt">
                                     <h8>PRK</h8></br></br>
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                                     <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="pcontent">
+
                                     <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>
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                                     <h8>CA (Carbonic Anhydrase)</h8></br></br>
 
                                     <p class="pcontent">
 
                                     <p class="pcontent">
 
                                         Achievements:</br>
 
                                         Achievements:</br>
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                                         the difference of XUI between two <i>E. coli</i> strain. We found out that both
 
                                         the difference of XUI between two <i>E. coli</i> strain. We found out that both
 
                                         strain
 
                                         strain
                                         shows similar trend: the XUI will be lower with the expression of the constructed protein. HoweverW3110 has a higher XUI compared with BL21 (DE3) in
+
                                         shows similar trend: the XUI will be lower with the expression of the constructed protein. However, W3110 has a higher XUI compared with BL21 (DE3) in
 
                                         constructed strain as well as the strain without plasmid. We infer two reasons
 
                                         constructed strain as well as the strain without plasmid. We infer two reasons
 
                                         that cause the difference of XUI:
 
                                         that cause the difference of XUI:
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                                     <ol>
 
                                     <ol>
                                         <li>W3110 “wildtype” strain has more flexible metabolic network but consumes
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                                         <li>W3110 “wildtype” strain has more flexible metabolic network. The carbon flux to pentose phosphate pathway of W3110 is more than that of BL21 (DE3) and thus consumes
 
                                             more xylose compare to lab strains such as BL21 (DE3).</li>
 
                                             more xylose compare to lab strains such as BL21 (DE3).</li>
  
 
                                         <li>The constructed protein expression in W3110 may be less than BL21 (DE3) lab
 
                                         <li>The constructed protein expression in W3110 may be less than BL21 (DE3) lab
                                             strain. BL21 (DE3) commonly used to express protein. We inferred that with
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                                             strain. BL21 (DE3) is commonly used to express protein. We inferred that with
 
                                             more protein been expressed, the bypass pathway in BL21 (DE3) will be more
 
                                             more protein been expressed, the bypass pathway in BL21 (DE3) will be more
 
                                             favored than the W3110 strain.</li>
 
                                             favored than the W3110 strain.</li>
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                                     <p class="pcontent">
 
                                     <p class="pcontent">
 
                                         Finally, we compare the XUI under different CO<sub>2</sub> concentration. We
 
                                         Finally, we compare the XUI under different CO<sub>2</sub> concentration. We
                                         incubated the
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                                         incubated the engineered
 
                                         bacteria in normal incubator without CO<sub>2</sub> input and the cell culture
 
                                         bacteria in normal incubator without CO<sub>2</sub> input and the cell culture
 
                                         incubator
 
                                         incubator
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                                         Fig 18. The comparison of the growth and the XUI of the BL21 (DE3) that contains
 
                                         Fig 18. The comparison of the growth and the XUI of the BL21 (DE3) that contains
 
                                         all three enzymes in normal incubator and 5% CO<sub>2</sub> incubator. The
 
                                         all three enzymes in normal incubator and 5% CO<sub>2</sub> incubator. The
                                         strain was grown in
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                                         strain grown in
 
                                         CO<sub>2</sub> incubator showed better growth and lower XUI, which indicates
 
                                         CO<sub>2</sub> incubator showed better growth and lower XUI, which indicates
 
                                         that our
 
                                         that our
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                                         <div id="pt">
 
                                         <div id="pt">
 
                                             <p class="pcontent">
 
                                             <p class="pcontent">
                                                     The engineered <i>E. coli</i> BL21 (DE3) are cultured in M9 medium with formula
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                                                     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 CO<sub>2</sub>
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                                                     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:
                                                    captured by
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                                                    the heterotrophic microbes, divide the net amount of carbon fixed by the mass
+
                                                    percent of carbon in biomass.
+
  
                                                    $${Ratio\ of\ carbon\ in\ CO_2\ fixed\ to\ carbon\ in\ biomass\ =\
+
 
                                                    {0.0069\over 0.0758}}$$
+
                                    </p>
                                                    $${=\ 9.1\%}$$
+
                                    <p class="pcontent">$${{{ \{ 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 Pasr will be induced at the pH value below four within 30 minutes. The
+
                                         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 Pasr is an acidic
+
                                         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 Pasr constructed pH sensing system can be used as an alert.
+
                                         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
                                         mild expression.
+
                                         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 fluorescent of previous and improved biobrick.</p>
+
                                         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

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