行政院國家科學委員會補助專題研究計畫成果報告:視網膜退化在犬模式的候選致病基因之分離、轉植、及表現
Date Issued
2000
Date
2000
Author(s)
DOI
892313B002138
Abstract
Canine generalized progressive retinal
atrophies (gPRA) are a group of degenerative
retinal diseases that are a major cause of
hereditary blindness in a number of dog breeds.
Expressed sequence tag (EST) approach was used
to identify and characterize potential candidate
genes from canine retinal cDNA libraries. A
conventional and subtractive canine retinal cDNA
libraries were constructed. For the conventional
cDNA library, the titer was 1.5 x 109 p.f.u./ml.
The average insert size was 1.6 kb and 96% of the
clones contained inserts. Differential hybridization
was performed to identify abundantly retinal
expressed cDNA clones. Random and
differentially expressed clones were fluorescently
labeled and sequenced. The sequences were
analyzed using GCG software and searched
against GenBank database. For genes of interest
isolated from the libraries, northern blotting and
RT-PCR were performed to determine mRNA
expression of the genes. DNA sequences from 76
differentially expressed clones and 100 random
cDNAs and analyzed. 54 out of 76 differentially
expressed cDNA clones (71%) showed homology
to known genes in the database. An unexpected
and interesting finding is the isolation of a
functionally important gene sFRP2, which is the
most abundant gene in the canine retina. The
remaining 22 cDNA clones (29%) showed no
homology to database sequences, representing
new sequences. In 100 random canine retinal
ESTs, 53 clones (53%) showed homology to
database sequences (including known genes and
ESTs). The remaining 47 cDNAs (47%) showed
no homology to database sequences. Four
candidate genes and 2 anonymous retinal ESTs
were selected to analyze mRNA expression. The
four known genes, namely opsin, cGMP-PDEA,
transducin 1A, and HRG4 showed retina-specific
expression. In anonymous ESTs, clone p61
revealed retina-specific expression, while p23
showed no tissue specificity. The isolation of
sFRP2 and HRG4 is the first finding in the canine
retina.
Subjects
retinal degeneration
retinal genes
canine genes
inherited disease
expressed
sequence tags
sequence tags
gene expression
Publisher
臺北市：國立臺灣大學獸醫學系暨研究所
Type
report
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