problems during protein expression

John A. Newitt newitt at removeme.nih.gov
Thu Jan 15 11:45:23 EST 1998


In article <34BD43B2.68BE at upei.ca>, jmorrison at upei.ca wrote:

> Hi.  My name is Jennifer Morrison.  I am a MSc student at the University
> of Prince Edward Island.  In our lab, we are expressing a de novo
> protein in bacterial cells and have had no problems in the past.  Lately
> our cells, when harvested, have been gray-black in color and the protein
> has not been showing up well, if at all, on gel.  If any of you happen
> to have experienced this type of problem or have any suggestions as to
> what the problem might be, please e-mail me at jmorrison at upei.ca

Are the cells gray-black in the culture or only after lysing? If only after
lysing, are you by chance using sonication? If so, the gray-black color
could be coming from the disintegration of your sonication probe. This
might also explain the lack of recovery of your protein since the
efficiency of sonication may also be less. Just a shot in the dark....

Regards,

John A. Newitt, Ph.D.           |   <newitt at removeme.nih.gov>
National Institutes of Health   |   FAX: 301-402-0387
Bethesda, Maryland  USA         |   

Please delete "removeme." from my address to reply by email.



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