GST-fusion yield 50mg/0.5 lt culture?

Virginia Urquidi vurquidi at molbiol.ox.ac.uk
Thu Oct 10 14:59:45 EST 1996


I'd appreciate the comments of those with experience in the subject.

The protein yields for GST fusion proteins according to the Pharmacia
purification protocols are the following:

                        50mg    10mg    1mg     50ug
Culture volume          20lt    4lt     20ml    1ml

Someone has just said to me that he produced 50mg of pure protein out of
half a liter culture. Is that feasible?

Also, the 0.5 lt culture was inoculated with the total of the 50ml overnight
culture (instead of the recommended 1:100 dilution). Is this sensible?  

Is the OD600 between 0.5 and 2 adequate for induction? What would happen if
you induce at much higher cell densities?

Thanks in advance for your comments





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*            V. Urquidi               *  vurquidi at molbiol.ox.ac.uk *
* Nuffield Dept Clinical Biochemistry *                            *
*     John Radcliffe Hospital         *   Tel.:+44(0)1865 221844   *
*    Headington, Oxford OX3 9DU       *   Fax: +44(0)1865 221834   *            
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