Wish to tag protein with SV40 T Ag NLS

Tom Thatcher ttha at uhura.cc.rochester.edu
Sat Dec 10 16:59:04 EST 1994


In <cotto-071294203535 at rmsentris.ls.nwu.edu> cotto at casbah () writes:

>Hi,
>I was wondering if there is an easy way to tag a protein of interest with
>the SV40 T antigen nuclear localization signal. Are there any vectors
>available for this purpose? I wish to avoid getting an oligo synthesized
>for generating the tagged protein. 

GOldfarb et al. (1986) Nature 322:641-644 describe how to chemically 
link an NLS peptide to a protein of interest.  If you want to do it
in the DNA, you can make your own DNA oligos, since the NLS is fairly
short.  Just make an oligo that encodes the NLS, with a restriction site
on each end along with extra spacer DNA so the restriction enzyme
will work.  Also make the complementary oligo.  Then just aneal them,
cut, and insert into a vector.

-- 
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
ttha at uhura.cc.rochester.edu           | to crack nuts.



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