I used lambda CE6, the phage that introduces the DE3 fragment into "non-DE3"
E.coli cells. I was comparing expression of a very toxic protein from pET
vector using the phage induction and the pLysS-amended approach. Got very
similar results in both cases, although there is more trouble involved with
the phage approach (plus the risk of contaminating the whole lab with the
lambda). My solution was not in finding the right molecular biology
contraption, but in selecting the more appropriate growth media. Thus, I got
5 times more protein on 2xYT + Mg then on plain LB. However, I still
preferred phage induction, because that allowed me to avoid extensive lysis
of cells before induction due to leakiness of the system and pLysS.
Emir
"Mark Bowen" <mboNOT at slac.stanford.edu> wrote in message
news:b146vm$2vk$1 at news.Stanford.EDU...
I am interested in learning more about phage induction of E. Coli.
Specifically introduction of T7 polymerase by phage. Does anyone have
any experience in this area?
--
Mark Bowen
