Expressed sequence tag analysis of human retina for the NEIBank Project: retbindin, an abundant, novel retinal cDNA and alternative splicing of other retina-preferred gene transcripts

Expressed sequence tag (EST) analysis was performed on un-normalized, unamplified cDNA libraries constructed from adult human retina to examine the expression profile of the tissue and to contribute resources for functional genomics studies. Two size fractionated cDNA libraries (designated hd and he...

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Bibliographic Details
Published in:Molecular vision 2002-06, Vol.8, p.196-204
Main Authors: Wistow, Graeme, Bernstein, Steven L, Wyatt, M Keith, Ray, Sugata, Behal, Amita, Touchman, Jeffrey W, Bouffard, Gerald, Smith, Don, Peterson, Katherine
Format: Article
Language:English
Subjects:
Aged
Aged, 80 and over
Alternative Splicing - genetics
Amino Acid Sequence
Base Sequence
Blotting, Northern
DNA, Complementary - analysis
Expressed Sequence Tags
Eye Proteins - genetics
Eye Proteins - metabolism
Gene Expression Profiling
Humans
Male
Molecular Sequence Data
National Institutes of Health (U.S.)
Oligonucleotide Array Sequence Analysis
Ophthalmology
Polymerase Chain Reaction
Retina - metabolism
United States
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Summary:Expressed sequence tag (EST) analysis was performed on un-normalized, unamplified cDNA libraries constructed from adult human retina to examine the expression profile of the tissue and to contribute resources for functional genomics studies. Two size fractionated cDNA libraries (designated hd and he) were constructed from human retina RNA. Clones were randomly selected for sequencing and analyzed using the bioinformatics program GRIST (GRouping and Identification of Sequence Tags). PCR, Northern blotting and other techniques have been used to examine selected novel transcripts. After informatics analysis, 2200 retina cDNAs yield 1254 unique clusters, potentially representing individual genes. Opsin is the most abundant transcript and other retina transcripts are prominently represented. One abundant cluster of cDNAs encodes retbindin, a novel, retina preferred transcript which has sequence similarity to riboflavin binding proteins and whose gene is on chromosome 19. Variant transcripts of known retina genes are also observed, including an alternative exon in the coding sequence of the transcription factor NRL and a skipped coding sequence exon in the phosphodiesterase gammasubunit (PDE6G). The new retina cDNA libraries compare favorably in quality with those already represented in public databases. They are rich in retina specific sequences and include abundant cDNAs for a novel protein, retbindin. The function of retbindin remains to be determined, but it is a candidate for flavinoid or carotenoid binding. Analysis of multiple clones for highly expressed retina genes reveals several alternative splice variants in both coding and noncoding sequences which may have functional significance. The validated set of retina cDNAs will contribute to a nonredundant set for microarray construction.
ISSN:1090-0535