disruption vs deletion

Morris F. Manolson mm6y+ at andrew.cmu.edu
Tue Nov 2 09:07:32 EST 1993


>often gene disruptions in yeast are made by simple insertions of markers
>into the open reading frame of a gene to be studied. I believe that
>this is dangerous and may result in wrong interpretations. Do you
>have examples in which such simple disruptions gave results different
>from clean deletions or replacements. And what were the consequences? I
>would greatly appreciate to have commments on this problem.

I don't have any stories about disruptions vs deletions but I know
of two cases were disruptions were made that accidently knocked out
two genes instead of the one intended.  Yeast genes are
packed in so tightly that if you do not know exactly 
where the start and stop of your gene is, you may easily disrupt not
one but two genes.  Robert Preston (Hi Rob!) was working on PEP3.  
If you look at the sequence (Mol. Cel. Biol.
Vol. 11, p.5801, 1991) you will note that there is a VERY small ORF
just 120 basepairs away from the start of PEP3.  Rob's first
disruption knocked out both genes resulting in a lethal phenotype.
As it turned out, it was the itty-bitty ORF that encoded an essential
gene.  PEP3, as expected was just required for vacuolar function.

Take care, Morrie




******************************************************************************
Morris F. Manolson                     Tel:  416-813-6662  (office)
Division of Cell Biology                     416-813-5594  (lab)
Hospital for Sick Children                   416-813-5028  (FAX)
88 Elm St., McMaster building        email:  Morrie at resunix.ri.sickkids.on.ca
Toronto, Ontario, Canada
M5G 1X8



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