Problems with expression in E.coli

Dr. Duncan Clark news at
Fri Mar 23 07:28:13 EST 2001

In article <3ABB23C6.C3E0E684 at>, the eminent Sergio at UCM
>several months of working with them and sending them from lab to lab, they
>didn't express at all (only the original clones worked).

Not too surprising. That is a good example of why one should always work
from master stocks and not passage cultures repetitively.

> I switched to MBP
>fusion system and expression (and specially solubility) are much better, and
>purification are simpler and needs less whasing steps (and no imidazol to worry

Same applies as above. Go back to a master -70C stock for every culture
otherwise you will select for the most non-toxic (metabolic load,
lethality or whatever to the cell) variants.

The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

Duncan Clark
GeneSys Ltd.
Tel: +44(0)1252376288
FAX: +44(0)8701640382

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