Auto-induction protein production


Omikron / Photo Researchers, Inc

The paper:
F.W. Studier, "Protein production by auto-induction in high-density shaking cultures," Protein Express Purif, 41:207-34, 2005. (Cited in 124 papers)

The finding:
While working on the National Institutes of Health's structural genomics project, William Studier at the Brookhaven National Laboratory devised a method that produced up to 10 times the protein yields of conventional techniques and induced simultaneous expression in Escherichia coli cells.

The method:
Studier grew the cells in sync by blocking the entrance of lactose into the cells with glucose until the cells reached a certain growth. Once cells consume the glucose, lactose enters the cells and allows induction of proteins. "It's a great timesaver," Studier says.

The fine-tuning:
"You can manipulate the medium and by doing that change expression level," says Brian Fox from the University of Wisconsin, who uses the auto-induction technique in his work on enzymes. "The original Studier medium is 0.2% weight-to-volume lactose, and about 0.2% to 0.3% weight-to-volume glycerol. We get the best expression at 0.6% lactose and around almost 1% glycerol."

The next step:
James Hartley, director of the protein expression laboratory at the National Cancer Institute, has expressed two dozen different proteins using auto-induction. He says he's working to design an instrument that can auto-induce temperature-dependent protein expression.


The numbers:
Lactose
concentration
Optical density/culture growth
(light absorption at 600 nm wavelength)
0.05% 4.2
0.02% 4.9
0.01% 5.5
0.005% 5.8



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Enhancing Protein Production
by Dr.Raam Shanthi

[Comment posted 2007-11-20 17:07:43]
What a boon to investigators who are struggling with purification of proteins expressed in very small amounts in normal cells!
Even if this technique works only in bacteria one could use genetic engineering to have the bacteria express any (mammalian protein for example) protein in large quantities. Just wonderful!






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