I am trying to induce pBAD promotor. The plasmid with this promotor was inserted into a wild type E. coli by electrotransformation. I tried several different concentration of arabinose but no sucess so far. The E. coli strain is fully sequenced and all the genes for arabinose and their CD are intact. Could it be that the bacteria uses all the arabinose which makes it less available for the promotor in the plasmid? any suggestions would be highly appreciated.
I have a low copy plasmid and I am using LB media for the growth.
thanks
Hi there,
Glucose has to be low in the medium. What medium do you use?
Are you sure of the construct you use?
Hello did you sequence the strain after the insertion to make sure what you are inducing is not mutated ? if you did then try inducing in M9 medium to eliminate the possibility that low level contaminating lactose from the tryptone in the LB is inhibiting the pBAD expression.
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