single cell protein concentration

James Bassuk bassuk at u.washington.edu
Thu Mar 20 18:21:39 EST 2003


On Wed, 19 Mar 2003, jennifer wrote:

> If i overexpress some protein, in the cell population, some cells may have a
> lot of that protein, some may have less.  Can I use western to distinguish
> between them?
> Or is there any method to measure a specific protein's concentration in a
> single bacterial cell?
>
> BTW, does anbody know that whether using IPTG as an inducer will cause a
> problem like that?
>

Can you please help me understand why you want to differentiate between
individual cells?  Recombinant cultures that respond to IPTG usually
involve a T7lac operon -- is this your system?

As long as you use antibiotics to guard against wild-type overgrowth, and
pay attention to good methodology (i.e. like picking a fresh colony to
grow up), then IPTG induction will result in all recombinant cells
responding in sync.

Jim




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