I have transformed E. coli BL21(DE3) with the recombinant pET28a(+) containing the target gene. I had induced the culture in LB broth with 0.05 mM IPTG at 18oC for 18 hours and over-expressed the protein. On SDS-PAGE an intense band is obtained. Now I want to induce the transformed cells with IPTG on LB-agar plate containing the substrate.
Can I induce the transformed E. coli cells with IPTG on LB-agar plates with substrate spread on it as I want to see the product formation in headspace by GC-MS analysis with the substrate (Farnesyl pyrophosphate) incorporated in the solid media containing induced cells. Will the same concentration of IPTG work?
one thing you can do. Grow your E.coli having recombinant gene. At OD 0.4, induce it with IPTG, incubate half to one hour, make dilution and then spread on ur LB agar with substrate plate...Hope it will work.
You could include IPTG on your solid media as well. A slightly lower concentration might be better here. You could also try autoinduction medium (see Studier, Protein Expr Purif 2005).
In my opinion is going to be better if you first grow your E.coli, induce it with IPTG an then you spread it on the agar. Remember that the protein induction is more efficient if you do it during the stationary growing phase.
