Optimization of Pichia Expression?

02 February 2014 1 8K Report

I am expressing a plant gene in Pichia and have observed a faint band of expected size on SDS-PAGE. However the yield is low (Approx 7 micro gram for 25mL culture volume). Please suggest me the process of optimization, What are the time points I should Check and the concentrations of Methanol for protein Induction.

What are the typical yields in Pichia for Plant proteins?

Galit Kuznets

Try to do the induction with MtOH every 12h instead of 24h.

You can repeat the transformation and select clones on higher selection (I used 1mg/ml Zeocin, produce about 5-10mg from 0.5L)

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