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大鼠胸腺中糖皮质激素反应性mRNA的克隆

Cloning of glucocorticoid-responsive mRNA in the rat thymus.

作者信息

Hoshikawa Y, Izawa M, Ichii S

机构信息

Division of Physiology, Tottori University School of Medicine, Yonago, Japan.

出版信息

Endocrinol Jpn. 1988 Jun;35(3):429-37. doi: 10.1507/endocrj1954.35.429.

Abstract

To elucidate the molecular mechanism of rat thymus involution induced by administration of glucocorticoid, we screened a cDNA library for polysomal poly(A)+RNA from adrenalectomized rat thymi by a differential colony hybridization method. Labeled cDNAs for mRNAs isolated from thymi of adrenalectomized rats and rats receiving dexamethasone (Dex) treatment were used as probes. Eight cDNA clones for mRNAs which had a diminished response to administration of Dex were isolated. The relative concentration of cloned mRNAs in the thymic polysomal RNA was significantly decreased 3 h after hormone administration, while changes in the nuclear RNA were not significant after Dex administration. One of the selected cDNA clones designated pRTGR-8 corresponded to mRNA of about 3,000 nucleotides, and a nuclear run-off transcription assay indicated that the rate of transcription of pRTGR-8 RNA was repressed by Dex administration. The cloned cDNAs obtained in this experiment may provide useful probes for studying the negative regulation mechanism of gene expression by glucocorticoids.

摘要

为阐明糖皮质激素诱导大鼠胸腺退化的分子机制,我们采用差异菌落杂交法,从肾上腺切除大鼠的胸腺中筛选了一个多核糖体多聚腺苷酸(poly(A)+)RNA的cDNA文库。以从肾上腺切除大鼠和接受地塞米松(Dex)治疗的大鼠胸腺中分离的mRNA的标记cDNA作为探针。分离出了8个对Dex给药反应减弱的mRNA的cDNA克隆。激素给药3小时后,胸腺多核糖体RNA中克隆mRNA的相对浓度显著降低,而Dex给药后核RNA的变化不显著。其中一个选定的cDNA克隆命名为pRTGR-8,对应约3000个核苷酸的mRNA,核转录分析表明Dex给药可抑制pRTGR-8 RNA的转录速率。本实验获得的克隆cDNA可能为研究糖皮质激素对基因表达的负调控机制提供有用的探针。

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