Team:British Columbia/Notebook/Week 12
From 2011.igem.org
(Created page with "{{Template:Notebook}}") |
Watermelon (Talk | contribs) |
||
Line 1: | Line 1: | ||
{{Template:Notebook}} | {{Template:Notebook}} | ||
+ | |||
+ | ===Characterization: Limonene Synthase=== | ||
+ | |||
+ | Daisy is starting the expression of the Limonene synthase (which produces +limonene) in C41 DE3. Daisy has been bugging Chris Keeling a lot about the expression conditions. It turns out that Daisy needs to do an in vitro assay for limonene synthase (adding GPP + purified enzyme). The only problem is there is no tag for a purification step. So it will most likely be the limonene synthase + a whole bunch of other enzymes + GPP (substrate needed to convert to limonene). | ||
+ | |||
+ | This is potentially problematic since it is much harder to do an in vitro assay without a purification step. Daisy originally planned to do a 500 mL expression, as suggested by Bohlmann Lab members, which is what they usually do. However, Daisy is now going to do a 2 L or 3 L expression. It will be better to have more enzyme rather than less for the in vitro assay. |
Revision as of 19:30, 21 August 2011
Characterization: Limonene Synthase
Daisy is starting the expression of the Limonene synthase (which produces +limonene) in C41 DE3. Daisy has been bugging Chris Keeling a lot about the expression conditions. It turns out that Daisy needs to do an in vitro assay for limonene synthase (adding GPP + purified enzyme). The only problem is there is no tag for a purification step. So it will most likely be the limonene synthase + a whole bunch of other enzymes + GPP (substrate needed to convert to limonene).
This is potentially problematic since it is much harder to do an in vitro assay without a purification step. Daisy originally planned to do a 500 mL expression, as suggested by Bohlmann Lab members, which is what they usually do. However, Daisy is now going to do a 2 L or 3 L expression. It will be better to have more enzyme rather than less for the in vitro assay.