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Growing cells in presence of substrate

protein expression phosphate

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#1 Kmu

Kmu

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Posted 25 September 2013 - 11:49 AM

Hello,

 

I am working on a protein complex which is activated by phosphate starvation conditions and ends up utilizing phosphonates or phosphites as phosphate sources in vivo. I have been able to purify the entire complex but it is not active. I was thinking of growing my cells (BL-21 DE3) in a phosphate starvation media supplementing it with a phosphonate source to see if that makes the complex active. I was wondering if anyone has any experience with making low or no phosphate media? Also, any suggestions about growing cells in the presence of substrates will be most welcome.

 

Thank you.







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