Hi Mark,
Proper co-expression in trans requires that you use plasmids each with
different origins of replication and antibiotic markers, such as p15 +
Kan and ColE1 + Amp. I think the pET vectors of the DUET system use
these two origins (p15 and ColE1). This ensures much more efficient
dual transformation and proper plasmid stability over time. The two
backbone constructions above are used in a variety of pET vectors, so
if you look for this detail, you can co-transform and co-express from
many available plasmids and customize as you like. Suppliers will
tell you the origins that are used in each of the plasmids they sell.
Cheers,
Brian
=========================
Brian L. Mark, MSc, PhD
Assistant Professor
Department of Microbiology
Room 418, Buller Building
University of Manitoba
Winnipeg, Manitoba
CANADA R3T 2N2
Phone (204) 480-1430
Fax (204) 474-7603
Web: http://www.umanitoba.ca/science/microbiology/staff/mark/
On 2-Apr-08, at 8:55 AM, Mark J. van Raaij wrote:
Dear All,
Anyone have experience with the NovaGen Duet co-expression vectors?
Or can recommend others?
http://www.emdbiosciences.com/html/NVG/Duet_Spot.html
Greetings,
Mark
Mark J. van Raaij
Dpto de Bioquímica, Facultad de Farmacia
Universidad de Santiago
15782 Santiago de Compostela
Spain
http://web.usc.es/~vanraaij/
