Team:Stockholm/30 June 2010

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Contents

Morning meeting

We discussed the project proceedings.

  • It was decided that already amplified genes (cloned in pEX vector) should be transferred to 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.

Andreas

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.