Antiobiotics for protein expression
Posted 04 March 2011 - 09:42 AM
I would like to ask a simple question in order to determine if someone of you has such an experience. I am trying to express a Human enzyme in E.coli and the gene sequence of this enzyme contains many rare codons for e.coli. For that reason, I selected the strain BL 21 DE3 Rosetta gami 2. I got expression , but the yield is enough low (1.5 mg/L culture). It is stated that this strain has resistance for both chloramphenicol and kanamycin also. My vector (pET 14b) contains ampicillin gene resistance.
My question is simply: should I use both antibiotics for rosetta strain, e.g. ampicillin & chloramphenicol or just 1 is enough? will it affect the expression yield, or not? Do you have such an experience?
Thank you very much for your reply in advance.
All the best,
Posted 04 March 2011 - 11:23 AM
Posted 06 March 2011 - 11:05 PM
Posted 07 March 2011 - 05:12 AM
If there is sth I could help you, just let me know.