We tried to get the bacteriocin from E. coli ER2566. To check whether the protein had been expressed, we used SDS-PAGE to confirm. Because our sequences content Intein and Chitin Binding Domain(CBD), which are 28 kDa, the result should be check as the bacteriocins’ initial mass plus 28 kDa. The figure showed our SDS-PAGE result. From here, we can know our target bacteriocin is produced.
Difference between revisions of "Team:NCTU Formosa/Wet Lab/Expression"
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| − | We tried to get the bacteriocin from E. coli ER2566. To check whether the protein had been expressed, we used SDS-PAGE to confirm. Because our sequences content Intein and Chitin Binding Domain(CBD), which are 28 kDa, the result should be check as the bacteriocins’ initial mass plus 28 kDa. The figure showed our SDS-PAGE result. From here, we can know our target bacteriocin is produced. | + | We tried to get the bacteriocin from <i>E. coli</i> ER2566. To check whether the protein had been expressed, we used SDS-PAGE to confirm. Because our sequences content Intein and Chitin Binding Domain(CBD), which are 28 kDa, the result should be check as the bacteriocins’ initial mass plus 28 kDa. The figure showed our SDS-PAGE result. From here, we can know our target bacteriocin is produced. |
</p> | </p> | ||
</div> | </div> | ||
| − | <img src="https://static.igem.org/mediawiki/2018/6/65/T--NCTU_Formosa--expression_96_b.png" class="expression" style="display:block; margin:auto;" | + | <img src="https://static.igem.org/mediawiki/2018/6/65/T--NCTU_Formosa--expression_96_b.png" class="expression" style="display:block; margin:auto;" width="700"> |
| − | + | ||
<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 result of the bacteriocin. <br><p>(N1) Negative control: <i>E. coli</i> ER2566 without plasmid (N2) Negative control: <i>E. coli</i> ER2566 with empty plasmid <br>(A) Enterocin 96+intein+CBD(35.9kDa) (B) Enteroicin B+intein+CBD(35.5kDa) </p> | |
| + | </div> | ||
| + | <img src="https://static.igem.org/mediawiki/2018/f/f7/T--NCTU_Formosa--expression_lu_dur.png" class="expression" style="display:block; margin:auto;" width="700"> | ||
| + | <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> | ||
| + | Figure 2: SDS-PAGE result of the bacteriocin. <br><p>(N1) Negative control: <i>E. coli</i> ER2566 without plasmid (N2) Negative control: <i>E. coli</i> ER2566 with empty plasmid <br>(C) Leucocyclicin Q+intein+CBD(34.4kDa) (D) Durancin +intein+CBD(35.3kDa) </p> | ||
</div> | </div> | ||
</div> | </div> | ||
Revision as of 16:27, 29 September 2018
Protein Expression
Bacteriocin |
Mass(kDa) |
|---|---|
| Subtilocin | 5.8 |
| Enterocin A | 6.9 |
| Enterocin B | 7.5 |
| Enterocin 96 | 7.9 |
| Bovicin HJ50 | 6.25 |
| Durancin | 7.3 |
| Leucocyclicin Q | 6.4 |
| Lacticin Z | 5.9 |
Figure 1: SDS-PAGE result of the bacteriocin.
(N1) Negative control: E. coli ER2566 without plasmid (N2) Negative control: E. coli ER2566 with empty plasmid
(A) Enterocin 96+intein+CBD(35.9kDa) (B) Enteroicin B+intein+CBD(35.5kDa)
Figure 2: SDS-PAGE result of the bacteriocin.
(N1) Negative control: E. coli ER2566 without plasmid (N2) Negative control: E. coli ER2566 with empty plasmid
(C) Leucocyclicin Q+intein+CBD(34.4kDa) (D) Durancin +intein+CBD(35.3kDa)
