INVITRO TRANSCRIPTION OF THE DROSOPHILA-ENGRAILED GENE
INVITRO TRANSCRIPTION OF THE DROSOPHILA-ENGRAILED GENE
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DOI：
10.1101/gad.2.1.68
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发表时间：
1988-01-01
影响因子：
10.5
通讯作者：
KORNBERG, T
中科院分区：
文献类型：
--
作者：
SOELLER, WC;POOLE, SJ;KORNBERG, T
从果蝇胚胎中制备了一种能够准确启动 engrailed 基因转录的酶系统。该系统已通过色谱分离成两个部分，这两个部分都是特定的转录所必需的。 DNA 酶足迹和竞争分析在这两个级分之一中检测到至少两个序列特异性 DNA 结合蛋白。这些蛋白质一起结合到转录起始位点 400 bp 内的八个区域。大多数含有这些结合位点的区域是动物体外转录所必需的。此外，起始位点下游和转录单元前 40 个残基内的区域对于转录至关重要。瞬时体内表达测定表明，在果蝇组织培养细胞中转录需要这些相同的上游和下游序列。
An enzyme system that accurately initiates transcription of the engrailed gene has been prepared from Drosophila embryos. The system has been separated chromatographically into two fractions, both of which are required for specific engrailed transcription. DNase footprint and competition analysis detected at least two sequence-specific DNA-binding proteins in one of these two fractions. Together, these proteins bind to eight regions within 400 bp of the transcription initiation sites. Most of the regions containing these binding sites are required for manimal engrailed transcription in vitro. In addition, a region downstream from the initiation sites and within the first 40 residues of the transcription unit is essential for transcription. Transient in vivo expression assays indicated that these same upstream and downstream sequences are required for tranacription in Drosophila tissue culture cells.