Difference between revisions of "Team:Hong Kong-CUHK/Description"
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<center><img src="https://static.igem.org/mediawiki/2015/3/31/Cuhk_partimprovementgenephoto3.jpg"></center> | <center><img src="https://static.igem.org/mediawiki/2015/3/31/Cuhk_partimprovementgenephoto3.jpg"></center> | ||
− | <p>Figure 1. The photo of 1% agarose gel electrophoresis. L: DNA ladder. Lane 1: PCR product of oprF encoding fromthe Azotobactervinelandii strain DJ genome.</p> | + | <p style="margin-bottom: 1.2em">Figure 1. The photo of 1% agarose gel electrophoresis. L: DNA ladder. Lane 1: PCR product of oprF encoding fromthe Azotobactervinelandii strain DJ genome.</p> |
<center><img src="https://static.igem.org/mediawiki/2015/5/5b/Cuhk_partimprovementgenephoto4.jpg"></center> | <center><img src="https://static.igem.org/mediawiki/2015/5/5b/Cuhk_partimprovementgenephoto4.jpg"></center> | ||
− | <p>Figure 2. Checking of recombinant plasmid using double digestion. L: DNA ladder. Lane 1-3: Recombination Template for pSB1C3-oprF(BBa_K1648045) with double digestion cut at EcoR1 and PstI sites, with single digestion at Pst1 site, without digestion.</p> | + | <p style="margin-bottom: 1.2em">Figure 2. Checking of recombinant plasmid using double digestion. L: DNA ladder. Lane 1-3: Recombination Template for pSB1C3-oprF(BBa_K1648045) with double digestion cut at EcoR1 and PstI sites, with single digestion at Pst1 site, without digestion.</p> |
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<center><img src="https://static.igem.org/mediawiki/2015/c/c6/Cuhk_partimprovementgenephoto2.jpg"></center> | <center><img src="https://static.igem.org/mediawiki/2015/c/c6/Cuhk_partimprovementgenephoto2.jpg"></center> | ||
− | <p>Figure 3. Checking of recombinant plasmid using double digestion. L: DNA ladder. Lane 1-3: Recombination Template for J13002-oprF(BBa_K1648048) with double digestion cut at EcoR1 and PstI sites, with single digestion at Pst1 site, without digestion.</p | + | <p style="margin-bottom: 1.2em">Figure 3. Checking of recombinant plasmid using double digestion. L: DNA ladder. Lane 1-3: Recombination Template for J13002-oprF(BBa_K1648048) with double digestion cut at EcoR1 and PstI sites, with single digestion at Pst1 site, without digestion.</p> |
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<center><img src="https://static.igem.org/mediawiki/2015/1/11/Cuhk_partimprovementgenephoto1.jpg"></center> | <center><img src="https://static.igem.org/mediawiki/2015/1/11/Cuhk_partimprovementgenephoto1.jpg"></center> | ||
− | Figure 4. Checking of recombinant plasmid using double digestion. L: DNA ladder. Lane 1-2: Recombination Template for R0040- | + | <p style="margin-bottom: 1.2em">Figure 4. Checking of recombinant plasmid using double digestion. L: DNA ladder. Lane 1-2: Recombination Template for R0040-oprF*(BBa_K1648049) with double digestion cut at EcoR1 and PstI sites, with single digestion at Pst1 site. |
− | oprF*(BBa_K1648049) with double digestion cut at EcoR1 and PstI sites, with single digestion at Pst1 site. | + | |
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<p><font face="Times New Roman" size="5pt">Experiment Set-up and Ongoing Test</p></font> | <p><font face="Times New Roman" size="5pt">Experiment Set-up and Ongoing Test</p></font> | ||
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<p style="margin-bottom: 1.5em">In the experiment, we will use the colour change of methylene blue as an indicator to compare the efficiency between the transformed bacteria with different oprF plasmids and wild type bacteria. The desired function of the oprF porin protein for this experiment is to allow the diffusion of (reduced) electron carriers in and out of the periplasmic membrane from the outside of the cell. As the electron carrier (e.g. NAD+) picks up an electron in the periplasmic space (i.e. being reduced to NADH) and diffuse out of the cell through the porin protein; the electron on the NADH will transfer to Methylene Blue (the mediator solution outside the cell). When the Methylene Blue gets reduced to form Leucomethylene Blue, it will have a colour change from the original blue colour to colourless. Hence, the rate of transmission of electron carrier is calculated by the rate of reduction of methylene blue. The experiment is planned to carry out soon. The plasmids will also be transformed into Azotobacter vinelandii for the construction of our MFC.</p></font> | <p style="margin-bottom: 1.5em">In the experiment, we will use the colour change of methylene blue as an indicator to compare the efficiency between the transformed bacteria with different oprF plasmids and wild type bacteria. The desired function of the oprF porin protein for this experiment is to allow the diffusion of (reduced) electron carriers in and out of the periplasmic membrane from the outside of the cell. As the electron carrier (e.g. NAD+) picks up an electron in the periplasmic space (i.e. being reduced to NADH) and diffuse out of the cell through the porin protein; the electron on the NADH will transfer to Methylene Blue (the mediator solution outside the cell). When the Methylene Blue gets reduced to form Leucomethylene Blue, it will have a colour change from the original blue colour to colourless. Hence, the rate of transmission of electron carrier is calculated by the rate of reduction of methylene blue. The experiment is planned to carry out soon. The plasmids will also be transformed into Azotobacter vinelandii for the construction of our MFC.</p></font> | ||
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Revision as of 22:55, 18 September 2015