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小鼠β-珠蛋白基因5'端转录起始复合物的组织特异性形成。

Tissue-specific formation of transcription-initiation complexes at the 5' end of the mouse beta major globin gene.

作者信息

Triadou P, Lelong J C, Gros F, Crepin M

出版信息

Eur J Biochem. 1983 Sep 1;135(1):163-9. doi: 10.1111/j.1432-1033.1983.tb07632.x.

Abstract

Accurate initiation of transcription in vitro requires, in addition to RNA polymerase II, factors present in soluble extracts of cultured cells. We have developed transcription system in vitro, which permits us to visualize the transcription-initiation complexes on a mouse beta globin restricted fragment from a recombinant beta globin bacteriophage DNA. Using the lambda fragments as internal controls this system has allowed us to assess the specificity of transcription with RNA polymerase II. Comparing extracts from cells and tissues expressing the globin genes (Friend cells, spleen and blood from phenylhydrazine-induced anemic mice) with those which do not (thymus, liver, PCC3 cells), we observed that specific initiation of transcription on the beta globin gene occurs only with soluble extracts from erythroid tissues. This tissue-specific transcription is partially sensitive to alpha-amanitin and occurs at the 5' end of the globin gene.

摘要

体外转录的准确起始除了需要RNA聚合酶II外,还需要培养细胞可溶性提取物中的一些因子。我们开发了一种体外转录系统,该系统使我们能够在来自重组β珠蛋白噬菌体DNA的小鼠β珠蛋白限制性片段上观察转录起始复合物。使用λ片段作为内部对照,该系统使我们能够评估RNA聚合酶II转录的特异性。将表达珠蛋白基因的细胞和组织(Friend细胞、苯肼诱导贫血小鼠的脾脏和血液)的提取物与不表达珠蛋白基因的细胞和组织(胸腺、肝脏、PCC3细胞)的提取物进行比较,我们观察到β珠蛋白基因上的特异性转录起始仅发生在红系组织的可溶性提取物中。这种组织特异性转录对α-鹅膏蕈碱部分敏感,且发生在珠蛋白基因的5'端。

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