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克隆的鸡δ-晶体蛋白基因在小鼠细胞中的组织特异性表达。

Tissue-specific expression of a cloned chick delta-crystallin gene in mouse cells.

作者信息

Kondoh H, Yasuda K, Okada T S

出版信息

Nature. 1983 Feb 3;301(5899):440-2. doi: 10.1038/301440a0.

Abstract

Crystallins are a group of soluble proteins specific to vertebrate lenses, and have been used successfully as molecular markers for studying lens differentiation. The synthesis and accumulation of delta-crystallin in the differentiating lens cells of chick, in particular, have been extensively studied (for review see refs 1-3). Recently, we have cloned a continuous stretch of a chick delta-crystallin gene which is specifically expressed in lens cells. We have injected this gene into nuclei of various mouse somatic cells. delta-Crystallin is totally absent from mammalian lenses in which it is replaced by gamma-crystallin, thus this xenogeneic injection system should allow us to study tissue-specific gene expression. We report here experiments which demonstrate that (1) the cloned delta-crystallin gene of chick is expressed in the mouse lens epithelial cells as efficiently as in the homologous chick cells; (2) delta-crystallin synthesized in the mouse lens epithelium has the native molecular weight, indicating correct splicing of the gene transcripts; (3) the expression is inefficient in the non-lens tissues examined, suggesting lens-specific expression of the delta-crystallin gene in the xenogenic environment. To our knowledge, this is the first demonstration that a cloned gene shows preferential expression in homologous cell types of different species.

摘要

晶状体蛋白是脊椎动物晶状体特有的一组可溶性蛋白质,已成功用作研究晶状体分化的分子标记。特别是鸡晶状体分化细胞中δ-晶状体蛋白的合成和积累已得到广泛研究(综述见参考文献1-3)。最近,我们克隆了一段鸡δ-晶状体蛋白基因的连续片段,该基因在晶状体细胞中特异性表达。我们已将该基因注射到各种小鼠体细胞的细胞核中。δ-晶状体蛋白在哺乳动物晶状体中完全不存在,在那里它被γ-晶状体蛋白所取代,因此这种异种注射系统应能让我们研究组织特异性基因表达。我们在此报告的实验表明:(1)克隆的鸡δ-晶状体蛋白基因在小鼠晶状体上皮细胞中的表达效率与在同源鸡细胞中的表达效率一样高;(2)在小鼠晶状体上皮细胞中合成的δ-晶状体蛋白具有天然分子量,表明基因转录本的剪接正确;(3)在所检测的非晶状体组织中表达效率低下,表明δ-晶状体蛋白基因在异种环境中具有晶状体特异性表达。据我们所知,这是首次证明克隆基因在不同物种的同源细胞类型中表现出优先表达。

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