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protein expression level and OD600 in E.coli

e.coli protein expression

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#1 Biogareth

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Posted 23 September 2011 - 01:26 AM

Hi all,

Recently when I do the MTP screening using E.coli as expression cells, and I stop the cell growth after 12 hours, because OD is acceptable and CV is quite low. But after incubation of 16 hours, the OD is also similar.
Now I am wondering the recombinant protein expression level is similar for both time, which time I have to choose? Maybe it's easy I just test the activity for both.
Before that, I hope I can have some suggestions from you.
Thanks!


Biogareth

#2 protolder

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Posted 25 September 2011 - 10:53 PM

hola, if your protein isn´toxic at level produced with choosen inducer concentration the culture will stay at stationary phase because any of the essential grow constituyent of the medium could be dissapear.In flask LB, for instance, can get about 3 OD600 units. I would harvest at the most longer time, or better I would analyze both samples to see if the relative amount of recombinant protein is the same or different. Buena suerte





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