Team:Bielefeld-CeBiTec/Notebook/Journal/CO2-fixation/Sep

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September









  • can_csoS1-4 respectively can_csoS1-4_csoS1D and sap
    • We tried to finish our carboxysome with and without csoS1D but we could not find a correct clone until the begining of october. We suggested that our colony PCR did not work but also with a restriction digestion we got no result for this construct.


  • csoS1-4_GFP and T7_sap

  • tkt
    • This week we wanted to purify the enzyme of tkt for the SBPase assay.
      • Cultivation of pet16b_tkt in 250 ml LB medium
      • Induction with 1 mM IPTG at OD 0.8. Taking samples:t0, t1, t2, t3, t15, t17
      • SDS-Page of cultivation result in correct bands at ~100kD
      • His-Tag purification of tkt
      • SDS-Page of His-Tag purification result in correct bands at imidazol concentration of ...

  • fba
    • This week we wanted to purify the enzyme of fba for the SBPase assay.
      • Cultivation of pet16b_fba in 250 ml LB medium
      • Induction with 1 mM IPTG at OD 0.8. Taking samples:t0, t1, t2, t3, t15, t17
      • SDS-Page of cultivation result in correct bands at ~40kD
      • His-Tag purification of fba
      • SDS-Page of His-Tag purification result in correct bands at imidazol concentration of ...



  • T7_sRNA:pfkA and ptac_sRNA:pfkA
    • We made a cultivation with our two constructs to see if there is a difference between induced, not induced and the wild type of E. coli. The cultivations were made in shaking flasks with a volume of 30 ml in 250 ml flasks at 37°C. The induction was with rhamnose for the T7 promotor and IPTG for the ptac. It lasted 13 hours. We made 2 biological and 2 technical replica for each culture. We took two samples (1 ml) of each biological replica for HPLC analysis that followed.