Cloning, characterization, and mRNA expression analysis of novel human fetal cochlear cDNAs

To identify novel genes that are expressed specifically or preferentially in the cochlea, we constructed a cDNA library enriched for human cochlear cDNAs using a suppression subtractive hybridization technique. We analyzed 2640 clones by sequencing and BLAST similarity searches. One hundred and fift...

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Bibliographic Details
Published in:Genomics (San Diego, Calif.) Calif.), 2003-10, Vol.82 (4), p.480-490
Main Authors: Luijendijk, M.W.J., van de Pol, T.J.R., van Duijnhoven, G., den Hollander, A.I., ten Caat, J., van Limpt, V., Brunner, H.G., Kremer, H., Cremers, F.P.M.
Format: Article
Language:English
Subjects:
Base Sequence
Biological and medical sciences
cDNA subtraction
Chromosome Mapping
Cloning, Molecular
Cochlea
Cochlea - metabolism
Deafness - genetics
DNA, Complementary - chemistry
DNA, Complementary - genetics
DNA, Complementary - metabolism
Fundamental and applied biological sciences. Psychology
Gene Expression
Gene Expression Profiling
Gene Library
Genes. Genome
Genetics of eukaryotes. Biological and molecular evolution
Hearing impairment
Humans
In Situ Hybridization - methods
Molecular and cellular biology
Molecular genetics
Molecular Sequence Data
mRNA expression profiling
Reverse Transcriptase Polymerase Chain Reaction
Sequence Analysis, DNA
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Summary:To identify novel genes that are expressed specifically or preferentially in the cochlea, we constructed a cDNA library enriched for human cochlear cDNAs using a suppression subtractive hybridization technique. We analyzed 2640 clones by sequencing and BLAST similarity searches. One hundred and fifty-five different cDNA fragments mapped in nonsyndromic hearing impairment loci for which the causative gene has not been cloned yet. Approximately 30% of the clones show no similarity to any known human gene or expressed sequence tag (EST). Clones mapping in nonsyndromic deafness loci and a selection of clones that represent novel ESTs were analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR) of RNA derived from 12 human fetal tissues. Our data suggest that a quarter of the novel genes in our library are preferentially expressed in fetal cochlea. These may play a physiologically important role in the hearing process and represent candidate genes for hereditary hearing impairment.
ISSN:0888-7543
1089-8646
DOI:10.1016/S0888-7543(03)00150-2