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| <div class="title"><p>Overview</p></div> | | <div class="title"><p>Overview</p></div> |
| <div class="text"> | | <div class="text"> |
− | <p>No matter how great our products, the most important thing that we need to concern is the safety of the users, producers, and the environment. Bacteriocins are natural peptides and degrade quickly in the environment. However, we need to confirm that the modified <i>E.coli</i> won’t spray into the environment, thus, we cut into 3 aspects to ensure the safety of our product: the training of the experimenter, the level and the rule of the laboratory, and the safety of our final products used in the environment.</p> | + | <p>Our system provides a nice Biosimulator, bacterioicn, which shows the antimicrobial activity against specific strains. Since we plan to utilize our product into reality, we have to make sure that these peptides will not cause any safety problems and our modified bacteria will not be spread into the environment.</p> |
| + | <p>Bacteriocins are natural peptides and degrade quickly in the environment. Therefore, the most important thing we need to confirm is the modified <i>E.coli</i>. Thus, we cut into different aspects to ensure the safety of our product: the training of the experimenter, the level and the rule of the laboratory, and the safety of our final products using in the environment.</p> |
| </div> | | </div> |
| </div> | | </div> |
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| <div class="title_1"><p>High-temperature Sterilization<br>Target on Modified <i>E.coli</i> ER2566</p></div> | | <div class="title_1"><p>High-temperature Sterilization<br>Target on Modified <i>E.coli</i> ER2566</p></div> |
| <div class="text"> | | <div class="text"> |
− | <p>Because our bacteriocins are heat stable, we use high-temperature sterilization to make sure our bacteriocins would not contain any alive <i>E. coli</i>. In order to prove our theory is right. We boiled our bacteriocins for 15, 30, and 45 minutes, and put them on LB Agar plate and cultured it at 37℃ for 16 hours. Compared with the sample which is not boiled, the figure shows the result of the plate and we can see that the boiled samples have no alive bacteria exist. | + | <p>Bacteriocins are usually heat stable, we use high-temperature sterilization to double make sure our peptide solution does not contain any living <i>E. coli</i>. However, peptides may degrades after long time sterilization. To find out the best fitted time for sterilization, we boiled our bacteriocins for 0, 15, 30, and 45 minutes, and put them on LB Agar plate and cultured it at 37℃ for 16 hours.</p> |
− | However, we still need to check whether our protein is degraded after boiling. We do SDS-PAGE to test the above samples. The result shows that the peptide doesn’t degrade after boiling for such a long time. Thus, we can produce the efficient products while preserving the safety of them. </p>
| + | <p>These are the result of the plates, we can easily observe that the bacteria exists only in the sample that is not boiled. After fifteen minutes of sterilization, there are no alive bacterias exist.</p> |
| </div> | | </div> |
− | <img src="https://static.igem.org/mediawiki/2018/7/71/T--NCTU_Formosa--Safety_96_B.png" class="sds"> | + | <img src="https://static.igem.org/mediawiki/2018/e/e6/T--NCTU_Formosa--96_safety_plate.png" class="sds_1"> |
| + | <img src="https://static.igem.org/mediawiki/2018/9/9e/T--NCTU_Formosa--B_safety_plate.png" class="sds_2"> |
| <div class="explanation"> | | <div class="explanation"> |
| <svg class="icon" aria-hidden="true" data-prefix="fas" data-icon="arrow-circle-up" class="svg-inline--fa fa-arrow-circle-up fa-w-16" role="img" xmlns="http://www.w3.org/2000/svg" viewBox="0 0 512 512"><path fill="currentColor" d="M8 256C8 119 119 8 256 8s248 111 248 248-111 248-248 248S8 393 8 256zm143.6 28.9l72.4-75.5V392c0 13.3 10.7 24 24 24h16c13.3 0 24-10.7 24-24V209.4l72.4 75.5c9.3 9.7 24.8 9.9 34.3.4l10.9-11c9.4-9.4 9.4-24.6 0-33.9L273 107.7c-9.4-9.4-24.6-9.4-33.9 0L106.3 240.4c-9.4 9.4-9.4 24.6 0 33.9l10.9 11c9.6 9.5 25.1 9.3 34.4-.4z"></path></svg> | | <svg class="icon" aria-hidden="true" data-prefix="fas" data-icon="arrow-circle-up" class="svg-inline--fa fa-arrow-circle-up fa-w-16" role="img" xmlns="http://www.w3.org/2000/svg" viewBox="0 0 512 512"><path fill="currentColor" d="M8 256C8 119 119 8 256 8s248 111 248 248-111 248-248 248S8 393 8 256zm143.6 28.9l72.4-75.5V392c0 13.3 10.7 24 24 24h16c13.3 0 24-10.7 24-24V209.4l72.4 75.5c9.3 9.7 24.8 9.9 34.3.4l10.9-11c9.4-9.4 9.4-24.6 0-33.9L273 107.7c-9.4-9.4-24.6-9.4-33.9 0L106.3 240.4c-9.4 9.4-9.4 24.6 0 33.9l10.9 11c9.6 9.5 25.1 9.3 34.4-.4z"></path></svg> |
− | Figure 1: SDS-PAGE gel and the concentrations of boiling test to bacteriocins.<p> (A) Enterocin 96+intein+CBD(35.9kDa) (B) Enteroicin B+intein+CBD(35.5kDa)</p> | + | Figure 5: LB Agar plate of boiling test to Enterocin 96+intein+CBD(35.9kDa) and Enteroicin B+intein+CBD(35.5kDa) |
| </div> | | </div> |
− | <img src="https://static.igem.org/mediawiki/2018/1/10/T--NCTU_Formosa--Safety_Bov_Dur.png" class="sds"> | + | <img src="https://static.igem.org/mediawiki/2018/a/a2/T--NCTU_Formosa--Bov_safety_plate.png" class="sds_1"> |
| + | <img src="https://static.igem.org/mediawiki/2018/3/33/T--NCTU_Formosa--Dur_safety_plate.png" class="sds_2"> |
| <div class="explanation"> | | <div class="explanation"> |
| <svg class="icon" aria-hidden="true" data-prefix="fas" data-icon="arrow-circle-up" class="svg-inline--fa fa-arrow-circle-up fa-w-16" role="img" xmlns="http://www.w3.org/2000/svg" viewBox="0 0 512 512"><path fill="currentColor" d="M8 256C8 119 119 8 256 8s248 111 248 248-111 248-248 248S8 393 8 256zm143.6 28.9l72.4-75.5V392c0 13.3 10.7 24 24 24h16c13.3 0 24-10.7 24-24V209.4l72.4 75.5c9.3 9.7 24.8 9.9 34.3.4l10.9-11c9.4-9.4 9.4-24.6 0-33.9L273 107.7c-9.4-9.4-24.6-9.4-33.9 0L106.3 240.4c-9.4 9.4-9.4 24.6 0 33.9l10.9 11c9.6 9.5 25.1 9.3 34.4-.4z"></path></svg> | | <svg class="icon" aria-hidden="true" data-prefix="fas" data-icon="arrow-circle-up" class="svg-inline--fa fa-arrow-circle-up fa-w-16" role="img" xmlns="http://www.w3.org/2000/svg" viewBox="0 0 512 512"><path fill="currentColor" d="M8 256C8 119 119 8 256 8s248 111 248 248-111 248-248 248S8 393 8 256zm143.6 28.9l72.4-75.5V392c0 13.3 10.7 24 24 24h16c13.3 0 24-10.7 24-24V209.4l72.4 75.5c9.3 9.7 24.8 9.9 34.3.4l10.9-11c9.4-9.4 9.4-24.6 0-33.9L273 107.7c-9.4-9.4-24.6-9.4-33.9 0L106.3 240.4c-9.4 9.4-9.4 24.6 0 33.9l10.9 11c9.6 9.5 25.1 9.3 34.4-.4z"></path></svg> |
− | Figure 2: SDS-PAGE gel and the concentrations of boiling test to bacteriocins.<p>(C)Bovicin HJ50+intein +CBD(34.25kDa) (D) Durancin +intein+CBD(35.3kDa)</p> | + | Figure 6: LB Agar plate of boiling test to Bovicin HJ50+intein +CBD(34.25kDa) and Durancin +intein+CBD(35.3kDa) |
| </div> | | </div> |
− | <img src="https://static.igem.org/mediawiki/2018/4/42/T--NCTU_Formosa--Safety_Lac.png" class="sds"> | + | <img src="https://static.igem.org/mediawiki/2018/1/12/T--NCTU_Formosa--Lac_safety_plate.png" class="sds_1"> |
| + | <img src="https://static.igem.org/mediawiki/2018/e/e3/T--NCTU_Formosa--Lu_safety_plate.png" class="sds_2"> |
| <div class="explanation"> | | <div class="explanation"> |
| <svg class="icon" aria-hidden="true" data-prefix="fas" data-icon="arrow-circle-up" class="svg-inline--fa fa-arrow-circle-up fa-w-16" role="img" xmlns="http://www.w3.org/2000/svg" viewBox="0 0 512 512"><path fill="currentColor" d="M8 256C8 119 119 8 256 8s248 111 248 248-111 248-248 248S8 393 8 256zm143.6 28.9l72.4-75.5V392c0 13.3 10.7 24 24 24h16c13.3 0 24-10.7 24-24V209.4l72.4 75.5c9.3 9.7 24.8 9.9 34.3.4l10.9-11c9.4-9.4 9.4-24.6 0-33.9L273 107.7c-9.4-9.4-24.6-9.4-33.9 0L106.3 240.4c-9.4 9.4-9.4 24.6 0 33.9l10.9 11c9.6 9.5 25.1 9.3 34.4-.4z"></path></svg> | | <svg class="icon" aria-hidden="true" data-prefix="fas" data-icon="arrow-circle-up" class="svg-inline--fa fa-arrow-circle-up fa-w-16" role="img" xmlns="http://www.w3.org/2000/svg" viewBox="0 0 512 512"><path fill="currentColor" d="M8 256C8 119 119 8 256 8s248 111 248 248-111 248-248 248S8 393 8 256zm143.6 28.9l72.4-75.5V392c0 13.3 10.7 24 24 24h16c13.3 0 24-10.7 24-24V209.4l72.4 75.5c9.3 9.7 24.8 9.9 34.3.4l10.9-11c9.4-9.4 9.4-24.6 0-33.9L273 107.7c-9.4-9.4-24.6-9.4-33.9 0L106.3 240.4c-9.4 9.4-9.4 24.6 0 33.9l10.9 11c9.6 9.5 25.1 9.3 34.4-.4z"></path></svg> |
− | Figure 3: SDS-PAGE gel and the concentrations of boiling test to bacteriocins.<p>(E) Lacticin+intein+CBD(33.9kDa)</p> | + | Figure 7: LB Agar plate of boiling test to Lacticin+intein+CBD(33.9kDa) and Leucocyclicin Q |
| </div> | | </div> |
| + | <img src="https://static.igem.org/mediawiki/2018/7/71/T--NCTU_Formosa--Safety_96_B.png" class="sds"> |
| <div class="explanation"> | | <div class="explanation"> |
| <svg class="icon" aria-hidden="true" data-prefix="fas" data-icon="arrow-circle-up" class="svg-inline--fa fa-arrow-circle-up fa-w-16" role="img" xmlns="http://www.w3.org/2000/svg" viewBox="0 0 512 512"><path fill="currentColor" d="M8 256C8 119 119 8 256 8s248 111 248 248-111 248-248 248S8 393 8 256zm143.6 28.9l72.4-75.5V392c0 13.3 10.7 24 24 24h16c13.3 0 24-10.7 24-24V209.4l72.4 75.5c9.3 9.7 24.8 9.9 34.3.4l10.9-11c9.4-9.4 9.4-24.6 0-33.9L273 107.7c-9.4-9.4-24.6-9.4-33.9 0L106.3 240.4c-9.4 9.4-9.4 24.6 0 33.9l10.9 11c9.6 9.5 25.1 9.3 34.4-.4z"></path></svg> | | <svg class="icon" aria-hidden="true" data-prefix="fas" data-icon="arrow-circle-up" class="svg-inline--fa fa-arrow-circle-up fa-w-16" role="img" xmlns="http://www.w3.org/2000/svg" viewBox="0 0 512 512"><path fill="currentColor" d="M8 256C8 119 119 8 256 8s248 111 248 248-111 248-248 248S8 393 8 256zm143.6 28.9l72.4-75.5V392c0 13.3 10.7 24 24 24h16c13.3 0 24-10.7 24-24V209.4l72.4 75.5c9.3 9.7 24.8 9.9 34.3.4l10.9-11c9.4-9.4 9.4-24.6 0-33.9L273 107.7c-9.4-9.4-24.6-9.4-33.9 0L106.3 240.4c-9.4 9.4-9.4 24.6 0 33.9l10.9 11c9.6 9.5 25.1 9.3 34.4-.4z"></path></svg> |
− | Figure 4: SDS-PAGE gel and the concentrations of boiling test to bacteriocins.<p>(F) Leucocyclicin Q+intein+CBD(34.4kDa)</p> | + | Figure 1: SDS-PAGE gel and the concentrations of boiling test to bacteriocins.<p> (A) Enterocin 96+intein+CBD(35.9kDa) (B) Enteroicin B+intein+CBD(35.5kDa)</p> |
| </div> | | </div> |
− | <img src="https://static.igem.org/mediawiki/2018/e/e6/T--NCTU_Formosa--96_safety_plate.png" class="sds_1"> | + | <img src="https://static.igem.org/mediawiki/2018/1/10/T--NCTU_Formosa--Safety_Bov_Dur.png" class="sds"> |
− | <img src="https://static.igem.org/mediawiki/2018/9/9e/T--NCTU_Formosa--B_safety_plate.png" class="sds_2">
| + | |
| <div class="explanation"> | | <div class="explanation"> |
| <svg class="icon" aria-hidden="true" data-prefix="fas" data-icon="arrow-circle-up" class="svg-inline--fa fa-arrow-circle-up fa-w-16" role="img" xmlns="http://www.w3.org/2000/svg" viewBox="0 0 512 512"><path fill="currentColor" d="M8 256C8 119 119 8 256 8s248 111 248 248-111 248-248 248S8 393 8 256zm143.6 28.9l72.4-75.5V392c0 13.3 10.7 24 24 24h16c13.3 0 24-10.7 24-24V209.4l72.4 75.5c9.3 9.7 24.8 9.9 34.3.4l10.9-11c9.4-9.4 9.4-24.6 0-33.9L273 107.7c-9.4-9.4-24.6-9.4-33.9 0L106.3 240.4c-9.4 9.4-9.4 24.6 0 33.9l10.9 11c9.6 9.5 25.1 9.3 34.4-.4z"></path></svg> | | <svg class="icon" aria-hidden="true" data-prefix="fas" data-icon="arrow-circle-up" class="svg-inline--fa fa-arrow-circle-up fa-w-16" role="img" xmlns="http://www.w3.org/2000/svg" viewBox="0 0 512 512"><path fill="currentColor" d="M8 256C8 119 119 8 256 8s248 111 248 248-111 248-248 248S8 393 8 256zm143.6 28.9l72.4-75.5V392c0 13.3 10.7 24 24 24h16c13.3 0 24-10.7 24-24V209.4l72.4 75.5c9.3 9.7 24.8 9.9 34.3.4l10.9-11c9.4-9.4 9.4-24.6 0-33.9L273 107.7c-9.4-9.4-24.6-9.4-33.9 0L106.3 240.4c-9.4 9.4-9.4 24.6 0 33.9l10.9 11c9.6 9.5 25.1 9.3 34.4-.4z"></path></svg> |
− | Figure 5: LB Agar plate of boiling test to Enterocin 96+intein+CBD(35.9kDa) and Enteroicin B+intein+CBD(35.5kDa) | + | Figure 2: SDS-PAGE gel and the concentrations of boiling test to bacteriocins.<p>(C)Bovicin HJ50+intein +CBD(34.25kDa) (D) Durancin +intein+CBD(35.3kDa)</p> |
| </div> | | </div> |
− | <img src="https://static.igem.org/mediawiki/2018/a/a2/T--NCTU_Formosa--Bov_safety_plate.png" class="sds_1"> | + | <img src="https://static.igem.org/mediawiki/2018/4/42/T--NCTU_Formosa--Safety_Lac.png" class="sds"> |
− | <img src="https://static.igem.org/mediawiki/2018/3/33/T--NCTU_Formosa--Dur_safety_plate.png" class="sds_2">
| + | |
| <div class="explanation"> | | <div class="explanation"> |
| <svg class="icon" aria-hidden="true" data-prefix="fas" data-icon="arrow-circle-up" class="svg-inline--fa fa-arrow-circle-up fa-w-16" role="img" xmlns="http://www.w3.org/2000/svg" viewBox="0 0 512 512"><path fill="currentColor" d="M8 256C8 119 119 8 256 8s248 111 248 248-111 248-248 248S8 393 8 256zm143.6 28.9l72.4-75.5V392c0 13.3 10.7 24 24 24h16c13.3 0 24-10.7 24-24V209.4l72.4 75.5c9.3 9.7 24.8 9.9 34.3.4l10.9-11c9.4-9.4 9.4-24.6 0-33.9L273 107.7c-9.4-9.4-24.6-9.4-33.9 0L106.3 240.4c-9.4 9.4-9.4 24.6 0 33.9l10.9 11c9.6 9.5 25.1 9.3 34.4-.4z"></path></svg> | | <svg class="icon" aria-hidden="true" data-prefix="fas" data-icon="arrow-circle-up" class="svg-inline--fa fa-arrow-circle-up fa-w-16" role="img" xmlns="http://www.w3.org/2000/svg" viewBox="0 0 512 512"><path fill="currentColor" d="M8 256C8 119 119 8 256 8s248 111 248 248-111 248-248 248S8 393 8 256zm143.6 28.9l72.4-75.5V392c0 13.3 10.7 24 24 24h16c13.3 0 24-10.7 24-24V209.4l72.4 75.5c9.3 9.7 24.8 9.9 34.3.4l10.9-11c9.4-9.4 9.4-24.6 0-33.9L273 107.7c-9.4-9.4-24.6-9.4-33.9 0L106.3 240.4c-9.4 9.4-9.4 24.6 0 33.9l10.9 11c9.6 9.5 25.1 9.3 34.4-.4z"></path></svg> |
− | Figure 6: LB Agar plate of boiling test to Bovicin HJ50+intein +CBD(34.25kDa) and Durancin +intein+CBD(35.3kDa) | + | Figure 3: SDS-PAGE gel and the concentrations of boiling test to bacteriocins.<p>(E) Lacticin+intein+CBD(33.9kDa)</p> |
| </div> | | </div> |
− | <img src="https://static.igem.org/mediawiki/2018/1/12/T--NCTU_Formosa--Lac_safety_plate.png" class="sds_1">
| |
− | <img src="https://static.igem.org/mediawiki/2018/e/e3/T--NCTU_Formosa--Lu_safety_plate.png" class="sds_2">
| |
| <div class="explanation"> | | <div class="explanation"> |
| <svg class="icon" aria-hidden="true" data-prefix="fas" data-icon="arrow-circle-up" class="svg-inline--fa fa-arrow-circle-up fa-w-16" role="img" xmlns="http://www.w3.org/2000/svg" viewBox="0 0 512 512"><path fill="currentColor" d="M8 256C8 119 119 8 256 8s248 111 248 248-111 248-248 248S8 393 8 256zm143.6 28.9l72.4-75.5V392c0 13.3 10.7 24 24 24h16c13.3 0 24-10.7 24-24V209.4l72.4 75.5c9.3 9.7 24.8 9.9 34.3.4l10.9-11c9.4-9.4 9.4-24.6 0-33.9L273 107.7c-9.4-9.4-24.6-9.4-33.9 0L106.3 240.4c-9.4 9.4-9.4 24.6 0 33.9l10.9 11c9.6 9.5 25.1 9.3 34.4-.4z"></path></svg> | | <svg class="icon" aria-hidden="true" data-prefix="fas" data-icon="arrow-circle-up" class="svg-inline--fa fa-arrow-circle-up fa-w-16" role="img" xmlns="http://www.w3.org/2000/svg" viewBox="0 0 512 512"><path fill="currentColor" d="M8 256C8 119 119 8 256 8s248 111 248 248-111 248-248 248S8 393 8 256zm143.6 28.9l72.4-75.5V392c0 13.3 10.7 24 24 24h16c13.3 0 24-10.7 24-24V209.4l72.4 75.5c9.3 9.7 24.8 9.9 34.3.4l10.9-11c9.4-9.4 9.4-24.6 0-33.9L273 107.7c-9.4-9.4-24.6-9.4-33.9 0L106.3 240.4c-9.4 9.4-9.4 24.6 0 33.9l10.9 11c9.6 9.5 25.1 9.3 34.4-.4z"></path></svg> |
− | Figure 7: LB Agar plate of boiling test to Lacticin+intein+CBD(33.9kDa) and Leucocyclicin Q | + | Figure 4: SDS-PAGE gel and the concentrations of boiling test to bacteriocins.<p>(F) Leucocyclicin Q+intein+CBD(34.4kDa)</p> |
| </div> | | </div> |
| </div> | | </div> |