Difference between revisions of "Team:Bordeaux/Results"

crdS, crdA and crdC genes ?

✵ crdS gene codes the Curdlan synthase.
✵ crdA gene codes a protein which assists translocation of nascent polymer across cytoplasmic membrane.
✵ crdC gene codes a protein which assists translocation of nascent polymer across the periplasm.
crdA, crdC, crdS genes occupy a contiguous 4,948-bp region in Agrobacterium sp. ATCC31749.

N.B : We tried to work on these three genes. However, amplification attempts by PCR were unsuccessful for crdA and crdC genes.So, in a first time, we focused on cloning crdS gene only.

OsmY promoter

In Agrobacterium sp.ATCC31749, Curdlan production is started after a nitrogen starvation in stationary phase. So we decided to use OsmY promoter (BBa_J45992) characterized by MIT 2006 iGEM team which is active in stationary phase and under high osmotic pressure condition. This promoter imitates Curdlan biosynthesis in E.coli without the nitrogen stress.

M63 and LB medium

Curdlan production was carried out in two different media: LB medium and M63 medium.
✵ We worked on M63 medium because this one is cited in literature. M63 is a minimal, low osmolarity medium for E.coli, resulting in slower growth rate of these cells. XXX Mettre la reference de la publication XXX →With this medium of known composition we were able to control parameters for the production of our molecule of interest.
✵We worked also on LB medium because this is the most common medium used in the laboratory.