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Isolation of genomic DNA fragments corresponding to genes modulated in vivo by a transcription factor.

机译:分离与转录因子在体内调节的基因相对应的基因组DNA片段。

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摘要

A new methodology for the identification of genes modulated by transcription factors in vivo is described. Mouse genomic DNA fragments bound by the thyroid hormone receptor (T3R) were selected and amplified in vitro. Subsequent hybridisation with biotinylated cDNA allowed the selection of those DNA fragments containing binding sites for T3R that corresponded to transcribed DNA. Expression analysis of the corresponding genes showed that more than 80% are indeed modulated by thyroid hormones in vivo in the liver. Together with the presence of consensus binding sites for T3R this result suggests that the selected DNA fragments may contain T3R transcriptional regulatory elements. This method, extensive to other ligand-modulated transcription factors, might be useful to all transcription factors with slight modifications.
机译:描述了一种在体内鉴定受转录因子调控的基因的新方法。选择并结合在甲状腺激素受体(T3R)上的小鼠基因组DNA片段。随后与生物素化的cDNA杂交,可以选择那些含有对应于转录的DNA的T3R结合位点的DNA片段。对相应基因的表达分析表明,肝脏中的体内甲状腺激素确实调节了80%以上。连同针对T3R的共有结合位点的存在,该结果表明所选的DNA片段可能包含T3R转录调控元件。该方法广泛应用于其他配体调节的转录因子,可能对所有经过轻微修饰的转录因子均有用。

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