Hello

I performed a digestion with 2 restriction enzymes to remove my insert (chimera of several antigenic sequences) from a cloning plasmid (PMA-RT), then cleaning and subsequent ligation with an expression vector (pET-22b) previously digested and isolated with the same restriction enzymes. At the time of transforming into E. coli BL21 (DE3)Plyss, they grow on the agar plate with antibiotics, but when I pass to the LB medium to obtain biomass, nothing grows. This stage is carried out in the presence of ampicillin (selection of pET- 22b).

Some comments talk about a possible toxicity of my insert for the bacteria E.coli B21(DE3) Plyss, but I don't know how to test this toxicity.

If I transform the PMA-RT + INSERT vector into E.colo BL21(DE3)PLyss, would you tell me if it is toxic to the bacteria?