nuclear localization/ mammalian vector

[Peter Horn]

Hello Netters,

We have a need for a mammalian expression vector, preferably SV40 or CMV
driven, that allows fusion of a protein of interest to a known nuclear
localization sequence (say of SV40 TAg for example). Has anyone out there
constructed such a vector for a positive control of nuclear localization?
Or is one available commercially? We have not run across one. Thanks for
any help you can provide.

Peter Horn

horn...@student.msu.edu

Michigan State University, Dept. of Biochemistry

[Tom Thatcher]

In <hornpete-2...@triezenberg2.bch.msu.edu> horn...@student.msu.edu (Peter Horn) writes:

>Hello Netters,

> We have a need for a mammalian expression vector, preferably SV40 or CMV
>driven, that allows fusion of a protein of interest to a known nuclear
>localization sequence (say of SV40 TAg for example). Has anyone out there
>constructed such a vector for a positive control of nuclear localization?
>Or is one available commercially? We have not run across one. Thanks for
>any help you can provide.

Look for the papers of David Goldfarb. He has done a lot of work on
nuclear localization. If anyone has a vector he does, or knows who has
it. One NLS he uses is a few amino acid peptide of SV40 large T--this has
the advantage that a single (defined) amino acid change renders it non-
functional. You could easily construct oligos to act as linkers
containing the + and - NLS and ligate them to your genes. The sequences
of the NLS are in his papers.

Or you could write to
Dr. David Goldfarb
Department of Biology
University of Rochester
Rochester, NY 14627

--
Tom Thatcher | You can give a PC to a Homo habilis,
University of Rochester Cancer Center | and he'll use it, but he'll use it
tt...@uhura.cc.rochester.edu | to crack nuts.