Selection of best recombinant protein produing strain - selection of industrial strain (Sep/16/2006 )
Hi there:
I'm working on several recombinant E. coli for production of a pharmacuetical proteins. I need to know wich factors are most important for selection of best strian and how kind of experimental design is needed? 
what strain depends on what protein you want to express, for example whether it is toxic or contains rare codons, there are a number of different e.coli systems which cater for these purposes.
also consider how you want to purify the protein, it may be insoluble and need some downstream processing to renature the enzyme after purification alternatively you may want to target expression to the periplasm or extracellular environment
there may also be some better systems out there, i have used a bacillus system that secreted my active protein into the medium, easy
just some things to think about before you choose.
i would get hold of a promega/novagen/invitrogen/amersham catalogue and have a look at each of the starins you are considering and decide which is right for you dependig on your selection criteria....
enjoy
[quote name='Jimmy_september' date='Sep 18 2006, 03:45 PM' post='69372']
what strain depends on what protein you want to express, for example whether it is toxic or contains rare codons, there are a number of different e.coli systems which cater for these purposes.
also consider how you want to purify the protein, it may be insoluble and need some downstream processing to renature the enzyme after purification alternatively you may want to target expression to the periplasm or extracellular environment
there may also be some better systems out there, i have used a bacillus system that secreted my active protein into the medium, easy
just some things to think about before you choose.
i would get hold of a promega/novagen/invitrogen/amersham catalogue and have a look at each of the starins you are considering and decide which is right for you dependig on your selection criteria....
Hi:
thanks but I'm working with five strain that have been cloned with rHGH and and my strains are bl21, TG1 & top10. the aim is periplasmic production of hGH and I need to know the best way for selection o thef best producer and efective factors in selection, and the next step is optimization of culture media and fermentation process for this porpose. 
If the process isn't already published, I can only say three words to you: small scale trials.