What makes a good mammalian 3'UTR?

John Ladasky ladasky at leland.Stanford.EDU
Mon Mar 24 16:59:16 EST 1997


Greetings,

	I've been working with a non-commercial eukaryotic expression vector
and I have been having trouble getting readable sequence.  I designed three
different primers that sit down in the region immediately 3' of the poly-
linker, and they all gave strong background bands.  After much haranguing, I
obtained an official sequence for this plasmid.  Surprise!  The 3' UTR that
is present next to the polylinker is also present downstream of the eukary-
otic neomycin resistance gene.  The duplicated region is nearly 300 base
pairs long!

	I am considering altering this plasmid so that the sequence next to
the polylinker appears only once, as it should.  I am wondering how hard it
is to change the 3' UTR for the neoR gene.  Is simply placing the AATAAA
sequence a few hundred bp downstream of the gene adequate?  Or are there 
other helpful signals?  Should I just grab a 3' UTR from another gene?


-- 
Unique ID : Ladasky, John Joseph Jr.
Title     : BA Biochemistry, U.C. Berkeley, 1989  (Ph.D. perhaps 1998???)
Location  : Stanford University, Dept. of Structural Biology
Keywords  : immunology, music, running, Green



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