E. coli MC4100 and T7 expression system - (Sep/05/2008 )
Hi there,
I need to use strain MC4100 of E. coli for overexpression of my favorite gene cloned into the pET28a vector.
I was wondering if I can use this strain the same way I use BL21 (DE3), ie, simply grow the transformed bacteria in LB + kanamycin and IPTG (to induce expression). In other words, does MC4100 has an IPTG-inducible T7 RNA polymerase encoding gene integrated in its chromosome ?
Thanks,
Seb
No, it has no T7 polymerase gene.
I need to use strain MC4100 of E. coli for overexpression of my favorite gene cloned into the pET28a vector.
I was wondering if I can use this strain the same way I use BL21 (DE3), ie, simply grow the transformed bacteria in LB + kanamycin and IPTG (to induce expression). In other words, does MC4100 has an IPTG-inducible T7 RNA polymerase encoding gene integrated in its chromosome ?
Thanks,
Seb
Why do you need to use this strain?
Actually I'd like to compare a metal reduction process between different engineered E. coli strains which membrane permeability is impaired. These mutants are derived from MC4100, so I need to use this one as a control. Some of these strains carry a pET vector in which a gene encoding a metal reductase was cloned. My preliminary experiments suggest that the IPTG induction may have not worked as expected. Before I check the protein production by western blot, I was just wondering if somebody had expertise in using the T7 expression system with MC4100 and derived strains. Thanks !