distronic expression plasmid

D P Dixon D.P.Dixon at durham.ac.uk
Thu Oct 5 03:40:22 EST 2000


In article <39DBFC86.5CC04183 at csb.yale.edu>,
Jeremy Murray  <jez at csb.yale.edu> wrote:
>wotsup
>
>Has anybody experience with over-expressing
>a distronic message using the same plasmid?
>If so I would be grateful if you could point 
>me to references, or if you would share your 
>strategy/experiences with the newsgroup.
>
>TIA. J.

Yes - good results expressing two proteins (to make a heterodimer) by
doubling up a pET-11d vector.
   i.e. T7 prom - RBS - cDNA1 - RBS - cDNA2 - terminator
See Plant Molecular Biology 40(6):997-1008 for details:
http://www.dur.ac.uk/~dbl1dpd/abstracts.htm#21

Expression levels in the dicistronic vector were comparable with those for
the corresponding monocistronic vectors.  I found that a poorly expressing
cDNA as the first cistron dragged down the expression of the second
cistron, but not vice versa.  Therefore, to get roughly equal expression
of the two cDNAs, put the best expresser last.
Hope this helps,

David Dixon






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