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oops - I just caught that the empty pET20 did transform well so the other suggestions about toxicity are probably more accurate. On Thu, 14 Jul 2011, Eric Larson wrote: > On Thu, 14 Jul 2011, Wonjin Bae wrote: > >> Hi, all >> >> Recently, some bactrial source enzyme(sialidase) was subcloned pET20b and >> pGEX4T3 vector. >> Then, these two construct were transformed to BL21(DE3) expression host. >> DNA sequencing results were accurate. >> >> In case of pGEX, many colony was formed and shown the high level of >> expression. >> But, colony was not shown in pET20b case. (no one colony) >> In case of mock-pET20a vector transformed very well. > > Simplest explanation - wrong antibiotic used on the pET20b case? > >> >> What's the problems? I never experieced transfomation problems. >> Does anyone know how to solve the this problems? >> >> Thanks a lot !! >> Genie, >> > > ================================ > Eric T. Larson, PhD > Biomolecular Structure Center > Department of Biochemistry > Box 357742 > University of Washington > Seattle, WA 98195 > > email: [log in to unmask] > ================================ >