IGEM:IMPERIAL/2006/project/popsblocker/Design: Difference between revisions
mNo edit summary |
No edit summary |
||
| Line 13: | Line 13: | ||
'''[[IGEM:IMPERIAL/Methodology/Part_documentation_template | Use this template]]''' | '''[[IGEM:IMPERIAL/Methodology/Part_documentation_template | Use this template]]''' | ||
==Cre-LoxP System== | ==Cre-LoxP System== | ||
==Registry== | |||
*insert part logo | |||
*insert link to the registry | |||
==Design choices== | |||
*demonstrate how the targeted specifications led to this specific design (bullet point style) | |||
*list how each specified inputs/outputs translate now in biological components | |||
*list possible sub-parts used in the design | |||
{| border="1" width="100%" | |||
|- style="background:lightgreen" | |||
! INPUTS !! biological component !! comments | |||
|- | |||
| input 1 || bla bla bla || bla bla bla | |||
|- | |||
| input 2 || bla bla bla || bla bla bla | |||
|- | |||
| input 3 || bla bla bla || bla bla bla | |||
|- style="background:orange" | |||
! OUTPUTS !! biological component !! comments | |||
|- | |||
| output 1 || bla bla bla || bla bla bla | |||
|- | |||
| output 2 || bla bla bla || bla bla bla | |||
|} | |||
==Open issues== | |||
*list known issues | |||
Revision as of 10:27, 22 October 2006
Cre-LoxP System
Registry
- insert part logo
- insert link to the registry
Design choices
- demonstrate how the targeted specifications led to this specific design (bullet point style)
- list how each specified inputs/outputs translate now in biological components
- list possible sub-parts used in the design
| INPUTS | biological component | comments |
|---|---|---|
| input 1 | bla bla bla | bla bla bla |
| input 2 | bla bla bla | bla bla bla |
| input 3 | bla bla bla | bla bla bla |
| OUTPUTS | biological component | comments |
| output 1 | bla bla bla | bla bla bla |
| output 2 | bla bla bla | bla bla bla |
Open issues
- list known issues
Design
Two devices are required:
- 1) A complementary sequence for a reporter with loxP sites either side
- 2) Cre recombinase under the control of a promoter
The device J37027 can be found here
Design of primers and method of construction
Proposed Implementation of Cre-LoxP system
Cre recombinase
Cre recombinase device in registry J37030
Sequence and primers here
There is one further step that we need to take if we want to successfully incorporate this system into the whole system. The plasmid containing the Cre enzyme coding region will be going into the prey cell. However the prey cell will already contain the main coding plasmid, which has ampicillin resistance. Therefore to ensure that the second plasmid is taken up into the cell as well, we need to swap plasmids by ligation. Ligating into a kanamycin resistant plasmid is probably our best bet. We could do this after the device has been constructed in the ampicillin resistant plasmid.
LoxP sites
The two LoxP sites that we are going to use are Lox66 and Lox71 - mutant LoxP sites
