Molecular cloning of a zinc finger gene eZNF from a human inner ear cDNA library, and in situ expression pattern of its mouse homologue in mouse inner ear.

A N Jacob, N A Manjunath, P Bray-Ward, R P Kandpal
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引用次数: 4

Abstract

We have isolated and characterized the cDNA for eZNF, a zinc finger gene expressed in human inner ear, from a kinetically enriched human inner ear cDNA library. The sequence of full length cDNA was determined and its expression pattern characterized. A high degree of homology is shared between eZNF and rat transcription factor Kid-1. It belongs to the C2H2 class of zinc finger genes, contains a Kruppel-associated box (KRAB) domain near the N-terminus, and has consensus sites for phosphorylation. The gene is expressed in kidney and inner ear structures of mouse and human as determined by Northern blot analysis. In situ hybridization was used to demonstrate specific expression of the mouse eZNF homologue in epithelial layers of the saccule, semicircular canals, and the cochlea of newborn mice. The genomic clone corresponding to the cDNA was isolated and used for fluorescence in situ hybridization to localize it to human chromosome 5qter. The identification of genes expressed in human inner ear by representational difference analysis, their chromosomal location, and expression pattern of their homologues in developing mouse inner ear comprise a strategy that can potentially identify genes important in hearing and deafness.

人内耳锌指基因eZNF的克隆及其小鼠同源基因在小鼠内耳中的原位表达模式
我们从一个动态富集的人内耳cDNA文库中分离并鉴定了锌指基因eZNF的cDNA。测定了其全长cDNA序列,并对其表达模式进行了表征。eZNF与大鼠转录因子Kid-1具有高度同源性。它属于锌指基因的C2H2类,在n端附近含有一个Kruppel-associated box (KRAB)结构域,并且具有公认的磷酸化位点。该基因在小鼠和人的肾脏和内耳结构中均有表达。采用原位杂交技术证实小鼠eZNF同源物在新生小鼠囊、半规管和耳蜗上皮层中的特异性表达。分离得到cDNA对应的基因组克隆,利用荧光原位杂交技术将其定位于人类染色体5qter。通过代表性差异分析鉴定人类内耳中表达的基因,它们的染色体定位以及它们在发育中的小鼠内耳中的同源物的表达模式,构成了一种潜在的识别听力和耳聋重要基因的策略。
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