Copenhagen/10 August 2011

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Wednesday

The single A2 colony plated yesterday in pSB1C3 were all red today = no insert!

Transformation of B1 and A2 in pSB1C3 from yesterday. B1 had lot of colonies but red = no insert. A2 had a lot of colonies as well, but some were white! 8 colonies were picked out and replated and an O/N culture made.

Lab Work

8 A2 colonies (pSB1C3) were picked out and replated and an O/N culture made.

Membrane preparation of O/N culture of IPTG induced A1 expressing cells.

Made overnight cultures for A2 in expression vector (233.3 and 233.4)

Measured absorption spectra on proteins from the membrane fraction. Despite help from Johan, Thomas, Peter and Birger, we did not succeed to confirm active CYP79A1. There might be a problem with the CO container or the dithionite used (either could be oxidated from the air, and therefore useless).

Other Work

Had a very nice meeting with Jaide from the DTU-2 Team. She gave us DNA pieces to do User Cloning on CYP79A2 and B1.

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