Title: Development and
characterization of a normalized canine retinal cDNA library for
genomic and expression studies
Zangerl B, Sun Q, Pillardy J, Johnson JL, Schweitzer PA,
Hernandez AG, Liu L, Acland GM, Aguirre GD
INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE 47 (6): 2632-2638
JUN 2006
Abstract: PURPOSE.
Identification of causative mutations for retinal blinding
disorders is often limited by restricted understanding of gene
expression and underlying molecular mechanisms that trigger
degenerative processes. This study was conducted to develop a
catalog of canine retina-expressed genes that would provide a
unique tool to investigate normal and altered function in the
adult retina. Because of the conserved syntenies between the dog
and human, this approach would identify new potential disease
candidate genes for both species.
METHODS. A canine normalized retinal cDNA library was
produced and analyzed by using a modified PhredPhrap algorithm.
Computerized annotation provided gene homology and chromosomal
location for individual clones and contigs in a Web-accessible
database.
RESULTS. From 6316 cDNA clones, 3980 retinal expressed
sequence tags (ESTs) were derived. Homology to the canine genome
draft sequence was found for more than 99% of all ESTs, but only
for 32% when compared with annotated canine cDNAs. Functional
analysis suggests an enrichment of this library for genes
involved with eye function and development, chaperone, or
ribosomal functions when compared with mouse and human National
Center for Biotechnology Information (NCBI) RefSeq entries.
CONCLUSIONS. A combination of annotation approaches with
ongoing mapping and expression studies provide functional data
covering at least 27% to 30% of the currently proposed canine
catalog of genes expressed in the retina. This is an essential
first step toward establishing an integrated network for gene
identification and expression patterns suitable for functional
genetics, comparative genomics and evolutionary analysis of
genes and gene families with respect to the developmental and
degenerative processes of the retina.
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