bacterial expression vector

Helen Niederwald helennie at my-deja.com
Thu Nov 30 08:39:43 EST 2000


In article <905g3n$2ced$1 at news.adamastor.ac.za>,
  "Johan Burger" <jtb at maties.sun.ac.za> wrote:
> Hi
>
> We need to express a eukaryotic (plant) gene in bacteria with the aim
to
> produce antibodies to the protein. Initial attempts (using the
Novagen pET
> 14b vector) were unsuccessful.

????
You may want to consider other Novagen vectors and/or strains. Some of
the pET vectors now carry lacI which will reduce expression under
uninduced conditions. Some BL21 derivatives are engineered to allow
fine-tuning of expression levels, so as not to get an all or nothing
response (Tuner strains). Others contain the pLys plasmid(s), also
reducing unwanted expression of toxic proteins. I would suggest to
consult their web page. They also have some info on using T7-RNA pol.
coming from a phage, rather than being on the chromosome.


Good luck,
Helen.




We expect that our protein might be toxic.
>
> Can anybody recommend a good prokaryotic system to express this
protein?
>
> Thanks
> Johan Burger
>
>


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