I want to express a GST fusion protein in pGEX-T2 vector. I have three E. coli strains and I don't know which one is best for the high expression of my fusion protein.
I have BL21 De3, Rosetta gami b De3 and BL21-plysS.
Rosetta strain is good if you want to express eukaryotic proteins, while pLysS help with toxic proteins. The best option is to transform your vector into all strains, do a test expression in small scale (50ml) in range of temperatures (16C, 18C ON, 37 for 3 h), different IPTG concentration and then choose optimal conditions to upscale. Load on gel samples before induction, after induction, soluble and pellet (after sonication). You will also know, if your protein is soluble. Good luck!
Rosetta strain is good if you want to express eukaryotic proteins, while pLysS help with toxic proteins. The best option is to transform your vector into all strains, do a test expression in small scale (50ml) in range of temperatures (16C, 18C ON, 37 for 3 h), different IPTG concentration and then choose optimal conditions to upscale. Load on gel samples before induction, after induction, soluble and pellet (after sonication). You will also know, if your protein is soluble. Good luck!
If you already have 3 strains, I would check them simultaneously. Will take you exactly the same time, like checking one. The result you will be able to verify on one gel. With the protein expression you usually don't know what exactly is not working, and what are the reasons for it. I wish you one kilogram of soluble protein;)
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