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黑腹果蝇唾液腺中蜕皮激素刺激的RNA合成:原位杂交检测

Ecdysone-stimulated RNA synthesis in salivary glands of drosophila melanogaster: assay by in situ hybridization.

作者信息

Bonner J J, Pardue M L

出版信息

Cell. 1977 Sep;12(1):219-25. doi: 10.1016/0092-8674(77)90199-4.

Abstract

3H-RNA was isolated from nuclei and cytoplasm of Drosophila melanogaster larval salivary glands after labeling in organ culture in the presence or absence of ecdysone. Hybridization to the sites of the ecdysone-induced puffs 74EF and 75B could be detected only if the RNA was labeled in the presence of ecdysone, while hybridization to the ecdysone-repressed puff 68C was found primarily in the cytoplasmic RNA sample labeled in the absence of ecdysone. Hybridization at the site of an ecdysone-insensitive puff, 50CD, was readily detectable in all RNA samples. RNA hybridizing to the unpuffed region 60D seemed to be restricted to the nucleus of salivary gland cells and was present at only a low level in cytoplasmic RNA samples. The data indicate that in situ hybridization can be used to detect specific hormone-induced changes in transcription.

摘要

在有或没有蜕皮激素存在的情况下,对黑腹果蝇幼虫唾液腺进行器官培养标记后,从其细胞核和细胞质中分离出3H-RNA。只有当RNA在蜕皮激素存在的情况下进行标记时,才能检测到与蜕皮激素诱导的胀泡74EF和75B位点的杂交,而与蜕皮激素抑制的胀泡68C的杂交主要在没有蜕皮激素的情况下标记的细胞质RNA样品中发现。在所有RNA样品中都很容易检测到在蜕皮激素不敏感的胀泡50CD位点的杂交。与未胀泡区域60D杂交的RNA似乎局限于唾液腺细胞的细胞核,并且在细胞质RNA样品中仅以低水平存在。数据表明,原位杂交可用于检测特定激素诱导的转录变化。

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