"
Team:Heidelberg/Toolbox Guide
From 2014.igem.org
(Difference between revisions)
| Line 230: | Line 230: | ||
Please save this link:<br /> | Please save this link:<br /> | ||
| - | <code>https://2014.igem.org/ | + | <code>https://2014.igem.org/<span id="pagename"></span>#<span data-bind="text: comeBackHash"></span></code> |
In order to generate your linker, <span data-bind="text: softwareName"></span> needs to know the scar amino acid sequence that is caused by circularization. In your case, it is <span data-bind="text: data.useSortase() ? 'LPETG' : data.exteins().N == 'XXX' ? 'just C' : (data.exteins().N + data.exteins().C)"></span>. | In order to generate your linker, <span data-bind="text: softwareName"></span> needs to know the scar amino acid sequence that is caused by circularization. In your case, it is <span data-bind="text: data.useSortase() ? 'LPETG' : data.exteins().N == 'XXX' ? 'just C' : (data.exteins().N + data.exteins().C)"></span>. | ||
Revision as of 06:07, 15 October 2014
use the intein
TOOLBOX to
1. Please go to www.rcsb.org and get a pdb file of your protein. If you cannot find one, we will not be able to assist you circularizing it.
Additionally, check the DNA sequence of your proteins for EcoRI, XbaI, SpeI, PstI and BsaI recognition sites. If there are E/X/S/P sites, you might have problems to change your backbone or add a promotor. If there is a BsaI recognition site, the cloning will be more difficult.
2. Do you want to use split inteins or sortase to circularize your protein?
- Successfully used in our project
- High efficiency
- In vivo circularization
- In vitro only
- Well-purified protein required
- Not successfully tested yet
3. Can your protein be easily expressed in E. coli?
4. Which exteins do you want to use? They will remain as scars in your circular protein.
. If you want to save time, check manually whether the ends are close together (approx. Å or closer). For example, you can use the Swiss-PdbViewer or PyMOL.
Please use to generate a linker for your circular protein. [LINK]
This step might take up to 11 days.
NILS – hier könnte dein instruction-file-ersatz stehen
NILS – hier auch
NILS – hier immernoch
NILS – hier ebnfalls und auch gerne noch umfangreicher
Please save this link:
https://2014.igem.org/#
In order to generate your linker, needs to know the scar amino acid sequence that is caused by circularization. In your case, it is .
