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]
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