nmt expression

S L Forsburg nospamforsburg at salk.edu
Mon Oct 11 21:12:12 EST 1999



> nmt1 leakiness?
> 
> Nicholas Owen (nicholas.owen at anat.ox.ac.uk)
> Fri, 8 Oct 1999 12:30:17 +0100
> 
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> Message-Id: <199910081305.GAA02305 at net.bio.net>
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> To: yeast at net.bio.net
> 
> Date: Fri, 8 Oct 1999 12:30:17 +0100
> 
> From: Nicholas Owen <nicholas.owen at anat.ox.ac.uk>
> 
> Subject: nmt1 leakiness?
> 
> Dear All,
> 
> I have heard comments that the nmt1 promoter leaks in the presence of
> thiamine, I just wondered if there are any publications out there that
> actually state this. I cannot remember seeing any, but then again I can
> easily miss it.
> 
> Thanks for your help

The nmt promoter is not completely off in the presence of thiamine as
attested by these assays:
1)  beta gal levels in reporter constructs are not zero;  for full
strength nmt, levels +thiamine are similar to those of attenuated nmt
-thia (Forsburg, (1993). Nucl. Acids Res. 21, 2955-2956)

2)  Complementation assays:  the only "shut off" constructs that work
are those involving quite unstable proteins (eg, cdc18; Kelly, T.J.,et
al (1993).  Cell 74:371-382), and not those involving more stable
constructs (eg., cdc19/mcm2; Forsburg, S.L., (1997) Genetics 147:1025-1041)


-- 
-susan
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