Date: Sat, 7 Nov 1998 13:31:20 -0800 (PST)
From: Graham Dellaire <genestructure at yahoo.com>
Subject: RE: TetR fusion vector
To: genstruc at net.bio.net
Hello,
I think your best bet is to make your TetR sequence by ordering two
oligos corresponding to the (-) and (+) strands and include some extra
bases for a common restriction enzyme so you can clone it afterwards.
So the idea is to anneal these two oligos together to make your
double-stranded linker which you clone into your expression vector or
gene targeting vector.
example BamHI-TetR-BamHI
If you need two three copies of TetR, you can either put them directly
in the linker or... and this is much easier is just select clones that
have ligated several copies together when you clone the linker into
your expression vector (you will have to probably sequence to know
exact number of copies or do partial BamHI digests).
As for selection it will depend on your system. If you tell me more I
can provide more info.
Cheers,
Graham Dellaire
P.S. Many groups out there have these vectors, so just look up papers
where TetR vectors were used and write the corresponding author. They
are obliged by many journals to provide materials (including plasmids)
as part of the publication/copyright agreement. Be Bold!
==
Graham Dellaire, Ph.D.
Moderator
bionet.genome.gene-structure
