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
Department of Human Genetics, University Medical Center Nijmegen, PO Box 9101, 6500 HB Nijmegen, The Netherlands.
Genomics. 2003 Oct;82(4):480-90. doi: 10.1016/s0888-7543(03)00150-2.
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.
为了鉴定在耳蜗中特异性或优先表达的新基因,我们使用抑制性消减杂交技术构建了一个富含人耳蜗cDNA的cDNA文库。我们通过测序和BLAST相似性搜索分析了2640个克隆。155个不同的cDNA片段定位于非综合征性听力损失位点,其致病基因尚未克隆。大约30%的克隆与任何已知的人类基因或表达序列标签(EST)没有相似性。通过对来自12种人类胎儿组织的RNA进行逆转录-聚合酶链反应(RT-PCR),分析了定位于非综合征性耳聋位点的克隆以及代表新EST的一系列克隆。我们的数据表明,文库中四分之一的新基因在胎儿耳蜗中优先表达。这些基因可能在听力过程中发挥重要的生理作用,并代表遗传性听力损失的候选基因。