Nucleic Acids Research, Vol 24, Issue 2 289-294, Copyright © 1996 by Oxford University Press
H Suzuki, T Yaoi, J Kawai, A Hara, G Kuwajima and S Wantanabe
We have developed a new method, designated restriction landmark cDNA
scanning (RLCS), which displays many cDNA species quantitatively and
simultaneously as two-dimensional gel spots. In this method cDNA species of
uniform length were prepared for each mRNA species using restriction
enzymes. After the restriction enzyme sites were radiolabeled as landmarks,
the labeled fragments were subjected to high resolution two-dimensional gel
electrophoresis. In analyses of cDNA samples from adult mouse liver and
brain (cerebral cortex, cerebellum and brain stem) we detected
approximately 500 and >1000 discrete gel spots respectively of various
intensities at a time. The spot patterns of the three brain regions were
very similar, although not identical, but were quite different from the
pattern for the liver. RNA blot hybridization analysis using several cloned
spot DNAs as probes showed that differences in intensity of the spots among
RLCS profiles correlated well with expression levels of the corresponding
mRNA species in the brain regions. Because the spots and their intensities
reflect distinct mRNA species and their expression level respectively, the
RLCS is a novel cDNA display system which provides a great deal of
information and should be useful for systematic documentation of
differentially expressed genes.
ARTICLES
Restriction landmark cDNA scanning (RLCS): a novel cDNA display system using two-dimensional gel electrophoresis
Shionogi Institute for Medical Science, Osaka, Japan.
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