Oscarliu117 (Talk | contribs) |
Oscarliu117 (Talk | contribs) |
||
Line 19: | Line 19: | ||
<a class="list-group-item list-group-item-action" href="#Construction">Construction</a> | <a class="list-group-item list-group-item-action" href="#Construction">Construction</a> | ||
<a class="list-group-item list-group-item-action" href="#Total_solution">Total solution</a> | <a class="list-group-item list-group-item-action" href="#Total_solution">Total solution</a> | ||
− | <a class="list-group-item list-group-item-action" href="# | + | <a class="list-group-item list-group-item-action" href="#Carbon_fixation">Carbon fixation</a> |
− | + | ||
− | + | ||
<a class="list-group-item list-group-item-action" href="#"><i class="fa fa-arrow-up fa-1x" | <a class="list-group-item list-group-item-action" href="#"><i class="fa fa-arrow-up fa-1x" | ||
aria-hidden="true"></i></a> | aria-hidden="true"></i></a> | ||
Line 458: | Line 456: | ||
</p> | </p> | ||
− | + | <h8>XUI Comparison between BL21(DE3) and W3110</h8> | |
− | + | <p class="pcontent"> | |
+ | We then compare the XUI value between BL21(DE3) and W3110 constructed strain. | ||
+ | When we design our IDT sequence, we link the CA directly to the promoter PLacI, | ||
+ | so we could not transform CA construct into W3110 strain. We thus compare the | ||
+ | XUI of strains that only contains Rubisco and PRK (<a href="http://parts.igem.org/Part:BBa_K2762011" | ||
+ | style="color:#28ff28;">BBa_K2762011</a>) , | ||
+ | (<a href="http://parts.igem.org/Part:BBa_K2762007" style="color:#28ff28;">BBa_K2762007</a>). | ||
+ | We found out that both strain shows similar trend: the XUI will | ||
+ | be lower with the expression of the constructed protein. The growth condition | ||
+ | of both constructed strains is similar for the first 12 hours. We then compare | ||
+ | the difference of XUI between two E. coli strain. We found out that both 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 | ||
+ | constructed strain as well as the strain without plasmid. We infer two reasons | ||
+ | that cause the difference of XUI: | ||
+ | </p> | ||
+ | <ol> | ||
+ | <li>W3110 “wildtype” strain has more flexible metabolic network but consumes | ||
+ | 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 | |
− | + | strain. BL21(DE3) commonly used to express protein. We inferred that with | |
− | + | more protein been expressed, the bypass pathway in BL21(DE3) will be more | |
− | + | favored than the W3110 strain.</li> | |
− | + | </ol> | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | <img class="contentimg col-6" src=" "> | |
− | + | <img class="contentimg col-6" src=" "> | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | <p class="pcontent"> | |
+ | Fig. 14 Shows the growth and the XUI of BL21(DE3) and W3110 strains. | ||
+ | </p></br> | ||
+ | |||
+ | <p class="pcontent"> | ||
+ | We finally concluded that the efficiency of the bypass pathway in BL21(DE3) is | ||
+ | better than that in the W3110 strain. | ||
+ | </p> | ||
+ | |||
+ | <h8>Incubation under different CO2 concentration</h8> | ||
+ | |||
+ | <p class="pcontent"> | ||
+ | Finally, we compare the XUI under different CO2 concentration. We incubated the | ||
+ | bacteria in normal incubator without CO2 input and the cell culture incubator | ||
+ | that maintains 5% C02 concentration. We observed that the strain in 5% CO2 | ||
+ | incubator has lower the XUI. The supply of sufficient CO2 can increase the | ||
+ | efficiency of the bypass pathway and enhance the growth. We can concluded that | ||
+ | our constructed pathway can be utilize carbon dioxide as one of its carbon | ||
+ | source from this result. | ||
+ | </p> | ||
+ | |||
+ | <img class="contentimg col-6" src=" "> | ||
+ | |||
+ | <img class="contentimg col-6" src=" "> | ||
+ | |||
+ | <p class="pcontent"> | ||
+ | Fig. 15 The comparison of the growth and the XUI of the BL21(DE3) that contains | ||
+ | all three enzymes in normal incubator and 5% CO2 incubator. The strain grown in | ||
+ | CO2 incubator shows better growth and lower XUI, which indicates that our | ||
+ | strain can use CO2 as a carbon source in the presence of high CO2 level. | ||
+ | </p> | ||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
− | |||
</div> | </div> | ||
+ | |||
</div> | </div> | ||
− | <div id=" | + | |
+ | <div id="Carbon_fixation"> | ||
</br></br></br></br> | </br></br></br></br> | ||
− | <h3> | + | <h3>Estimation of the amount of the carbon fixation</h3> |
<div id="pt"> | <div id="pt"> | ||
− | <p class="pcontent"> | + | <p class="pcontent"> |
− | + | To find out how much and how efficient genetically engineered E. coli can fix | |
− | + | carbon dioxide, we use the material balance concept to evaluate the | |
− | + | heterotrophic CO2 fixation process. Consider a system composed of a single | |
− | + | component, the general material balance can be written as:</br></br> | |
+ | |||
+ | {Input to the system} – {Output to the system} = {Accumulation in the system}</br></br> | ||
+ | |||
+ | A system can be defined as an arbitrary portion of a process considered for | ||
+ | analysis, in which in this case, is an engineered carbon capturing E. coli. | ||
+ | </p> | ||
</div> | </div> | ||
− | |||
+ | <img class="contentimg" src=""> | ||
− | |||
− | |||
− | |||
<div id="pt"> | <div id="pt"> | ||
− | < | + | <p class="pcontent"> |
− | + | The engineered E. coli BL21(DE3) are cultured in M9 medium with formula | |
− | + | adjusted so that xylose is the sole carbon source. The aforementioned M9 Medium contains | |
− | + | 0.4% xylose and 1/1000 LB medium (the carbon consumed from LB medium can be | |
− | + | ignored). By applying the law of conservation of mass, which states that mass | |
− | + | may neither be created nor destroyed, the material balance for carbon in an | |
− | + | engineered E. coli may simply be written as | |
− | + | </p> | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | </ | + | |
− | + | ||
</div> | </div> | ||
+ | |||
+ | |||
</div> | </div> | ||
+ | |||
</div> | </div> |
Revision as of 15:42, 12 October 2018
Results