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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
• 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.