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首页> 外文期刊>Genomics >Cloning, characterization, and mRNA expression analysis of novel human fetal cochlear cDNAs.
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Cloning, characterization, and mRNA expression analysis of novel human fetal cochlear cDNAs.

机译：新型人类胎儿耳蜗cDNA的克隆，表征和mRNA表达分析。

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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.
机译：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。在尚未克隆致病基因的非综合征性听力障碍基因座中定位的155个不同cDNA片段。大约30％的克隆与任何已知的人类基因或表达的序列标签（EST）没有相似性。通过逆转录酶-聚合酶链反应（RT-PCR）对源自12个人类胎儿组织的RNA进行分析，分析了非综合征性耳聋基因座中的克隆作图以及代表新型EST的克隆选择。我们的数据表明，我们图书馆中的新基因中有四分之一在胎儿耳蜗中优先表达。这些可能在听力过程中发挥重要的生理作用，并代表遗传性听力障碍的候选基因。

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