Oscarliu117 (Talk | contribs) |
Oscarliu117 (Talk | contribs) |
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<div id="pt"> | <div id="pt"> | ||
<h8>PRK</h8> | <h8>PRK</h8> | ||
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Achievements:</br> | Achievements:</br> | ||
</p> | </p> | ||
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</ol> | </ol> | ||
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We constructed PRK fragments from IDT DNA synthesis. After PCR amplification, | We constructed PRK fragments from IDT DNA synthesis. After PCR amplification, | ||
PRK is then cloned into pSB1C3 and transformed into DH5α. SDS-PAGE ensured that | PRK is then cloned into pSB1C3 and transformed into DH5α. SDS-PAGE ensured that | ||
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<div id="pt"> | <div id="pt"> | ||
<h8>CA</h8> | <h8>CA</h8> | ||
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Achievements:</br> | Achievements:</br> | ||
</p> | </p> | ||
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</ol> | </ol> | ||
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We cloned the DNA fragments into pSB1C3 plasmid after the gene is amplified | We cloned the DNA fragments into pSB1C3 plasmid after the gene is amplified | ||
with PCR. We transform the plasmid into DH5-alpha and BL21(DE3). Next, we | with PCR. We transform the plasmid into DH5-alpha and BL21(DE3). Next, we | ||
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<img class="contentimg" src=" "> | <img class="contentimg" src=" "> | ||
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Fig. 5 Confirmation of ca digestion</br> | Fig. 5 Confirmation of ca digestion</br> | ||
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<div id="pt"> | <div id="pt"> | ||
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We then ran the activity test of CA. In our bypass pathway, the function of CA | We then ran the activity test of CA. In our bypass pathway, the function of CA | ||
is to | is to | ||
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<div id="pt"> | <div id="pt"> | ||
<h8>Rubisco</h8> | <h8>Rubisco</h8> | ||
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Achievements:</br> | Achievements:</br> | ||
</p> | </p> | ||
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</ol> | </ol> | ||
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We constructed PRK fragments from IDT DNA synthesis. After PCR amplification of | We constructed PRK fragments from IDT DNA synthesis. After PCR amplification of | ||
the three subunits, rubisco is then cloned into pSB1C3 and transformed into | the three subunits, rubisco is then cloned into pSB1C3 and transformed into | ||
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<img class="contentimg" src=" "> | <img class="contentimg" src=" "> | ||
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Fig. 7 Confirmation of rbcL digestion.</br> | Fig. 7 Confirmation of rbcL digestion.</br> | ||
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</p> | </p> | ||
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After mining a lot of information from the publications, we found out a method | After mining a lot of information from the publications, we found out a method | ||
to determine the activity of rubisco by thin-layer chromatographic has been | to determine the activity of rubisco by thin-layer chromatographic has been | ||
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<div id="pt"> | <div id="pt"> | ||
<h8>Rubisco</h8> | <h8>Rubisco</h8> | ||
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Achievements:</br> | Achievements:</br> | ||
</p> | </p> | ||
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<h8>Overview</h8> | <h8>Overview</h8> | ||
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In the total solution experiment, we strive to measure the carbon fixation | In the total solution experiment, we strive to measure the carbon fixation | ||
amount of each sample. After reading numerous publications, we found out that | amount of each sample. After reading numerous publications, we found out that | ||
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</p> | </p> | ||
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The test samples below were incubated in an altered M9 medium which substitute | The test samples below were incubated in an altered M9 medium which substitute | ||
glucose to xylose. 1/1000 of LB medium was added to support some rare elements. | glucose to xylose. 1/1000 of LB medium was added to support some rare elements. | ||
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</p> | </p> | ||
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We defined a new index, Xylose Utilization Index, to describe the potential of | We defined a new index, Xylose Utilization Index, to describe the potential of | ||
carbon fixation. We can compare this index of each strain to find out the | carbon fixation. We can compare this index of each strain to find out the | ||
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</p> | </p> | ||
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To define the XUI index, we firstly made two assumptions: | To define the XUI index, we firstly made two assumptions: | ||
</p> | </p> | ||
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<img class="contentimg" src=" "> | <img class="contentimg" src=" "> | ||
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Fig. 9 shows the dry cell weight of BL21(DE3) incubated in altered M9 xylose | Fig. 9 shows the dry cell weight of BL21(DE3) incubated in altered M9 xylose | ||
medium. | medium. | ||
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</ol> | </ol> | ||
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After these two assumptions, the Xylose Consumption Index is designed to | After these two assumptions, the Xylose Consumption Index is designed to | ||
evaluate | evaluate | ||
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<img class="contentimg" src=" "> | <img class="contentimg" src=" "> | ||
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We use the Dinitrosalicylic Acid (DNS) reducing sugar assay to measure the | We use the Dinitrosalicylic Acid (DNS) reducing sugar assay to measure the | ||
xylose | xylose | ||
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<img class="contentimg" src=" "> | <img class="contentimg" src=" "> | ||
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Fig. 10 Shows the calibration line of DNS assay kit. | Fig. 10 Shows the calibration line of DNS assay kit. | ||
</p> | </p> | ||
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Before measuring the XUI, we observe the growth curve of each strain. We found | Before measuring the XUI, we observe the growth curve of each strain. We found | ||
out | out | ||
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<img class="contentimg" src=" "> | <img class="contentimg" src=" "> | ||
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Fig. 11 shows the growth of W3110(L5T7), BL21(DE3), W3110 incubated in normal | Fig. 11 shows the growth of W3110(L5T7), BL21(DE3), W3110 incubated in normal | ||
incubator for 24 hours. The growth of W3110(L5T7) is not obvious while other | incubator for 24 hours. The growth of W3110(L5T7) is not obvious while other | ||
strains shows growth after 24hours. | strains shows growth after 24hours. | ||
</p> | </p> | ||
− | + | ||
</div> | </div> | ||
Revision as of 13:53, 12 October 2018
Results