IGEM:Caltech/2008/Project/Lactose intolerance: Difference between revisions
| Line 1: | Line 1: | ||
=Lactose Summary= | =Lactose Summary= | ||
We | We want to make two plasmids:<br> | ||
Plasmid #1 - A constitutive promoter with <i>LacZ</i> and <i>LacY</i>.<br> | Plasmid #1 - A constitutive promoter with <i>LacZ</i> and <i>LacY</i>.<br> | ||
Plasmid #2 - A lactose inducible promoter with lysozyme and holin.<br> | Plasmid #2 - A lactose inducible promoter with lysozyme and holin.<br> | ||
Revision as of 14:50, 19 June 2008
Lactose Summary
We want to make two plasmids:
Plasmid #1 - A constitutive promoter with LacZ and LacY.
Plasmid #2 - A lactose inducible promoter with lysozyme and holin.
The general idea is to overexpress LacY (Lactose receptor protein) with LacZ (Beta-Galactosidase). When lactose is present, the cell will uptake lactose and will flip on the LacI / LacIQ promoter when enough lactose is present. We then express lysozyme and holin which will lyse the cell releasing beta-gal cleaving lactose into glucose and galactose. Hopefully, glucose and galactose will be taken up by the host in the colon.
Lactose Outline
6.19.08
We have decided to use the I732017 LacZ (1009/11H) because we could not transform I732005 LacZ (1018/6F).
We ordered primers for LacY, and we will need to PCR out LacY. We then need to run cite directed mutagensis on LacY by added 2H to prevent EIIA(GLUC) binding.
