User:AndreasConstantinou/30 June 2010
From 2010.igem.org
Morning meeting
We discussed the project proceedings.
- It was decided that already amplified genes (cloned in pEX vector) should be transferred to [http://partsregistry.org/Part:pSB1C3 pSB1C3] vector.
- IgG protease
- Superoxidase dismutase (SOD)
- yCCS
- bFGF
- Primers for not-yet amplified genes should be redesigned to include the complete Assembly standard 25 prefix and suffix.
- CPP
- Transportan 10
- LMWP
- TAT
- MITF
- Protein A Z-domain
- Tyrosinase
- Vitamin B9 genes
- CPP
- BL21(DE3) was chosen as our strain for IPTG-induced protein expression from pEX vector.
Expression of SOD and yCCS from pEX expression vector
Continued from 29/6
We realized that Top10 and DH5alpha are not suitable for IPTG-induced protein expression. SOD and yCCS expression was therefore not proceeded from ON cultures. Glycerol stocks were prepared from ON culture and pEX*SOD and pEX*yCCS plasmids were prepared.
Preparation of competent BL21(DE3) cells
ON culture was set and grown in 37°C ON with 250 rpm rotary shaking.