e coli strain selection for protein expression - (Jun/23/2010 )
Hello, in our lab we have BL21 pRARE chemically competent cells which I have been told are used in protein expression experiments. Does this mean the pRARE plasmid is already transformed into the strain? If so, how can I then transform in my plasmid expressing my protein? The pRARE gives chloramphenicol resistance while my plasmid gives kanamycin resistance. Should I grow the cells on plates with both or just kanamycin?
Thank you
pRARE has a p15a origin of replication and is therefore compatible with common expression vectors like the pET family who have a ColE1-like origin of replication ...so you don't have to worry!
I would use both antibiotics to prevent plasmid loss of both plasmids!
Good luck for your expression experiments!
Regards,
p
You'll need both antibiotics. You may need chlor in the recovery SOC medium, tho'.
When all else fails, I'd just check with the website...