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protein expression - (Oct/19/2005 )

Hi all, I have very dumb question regarding the overexpression of protein. If I use a wrong antibiotics, can it affect on the purification? for example, If I am supposed to use kanamycin but misuse ampcillin, what kind result can I get? Could you let me know about that, please?

Thank you.

-klee-

um...I would say you will get very little protein

if you use the wrong selection, any cells you get will probably not have your expression vector

-aimikins-

Thank you so much for your message. In fact, I checked the vector and I sitll have the right vector and it produced very small amount of protein--I checked wiht histag antibody. But, when I inserted this vector to a new clean cell, the cell produced only inculsion body of the target protein. Could you explain this phenomenon, please?

-klee-

QUOTE (klee @ Oct 19 2005, 06:15 PM)
Thank you so much for your message. In fact, I checked the vector and I sitll have the right vector and it produced very small amount of protein--I checked wiht histag antibody. But, when I inserted this vector to a new clean cell, the cell produced only inculsion body of the target protein. Could you explain this phenomenon, please?

From what I know, usually overexpression of protein in cells would lead to inclusion body... perhaps this time round, they manage to survive better, able to produce more of your protein.

-ecarg-