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Analysis of consensus sequence patterns in Giardia cytoskeleton gene promoters.

机译:贾第虫细胞骨架基因启动子的共有序列模式分析。

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摘要

Protein-coding genes in the ancient eukaryote Giardia lamblia lack typical promoter consensus elements. We have analysed the immediate 5' flanking sequences of seven genes of related function (structural cytoskeleton proteins) to identify shared DNA motifs that might have a role in transcription initiation. Transcription start sites for five genes have been determined previously. Genomic mapping and mRNA primer extension experiments demonstrate additionally that the genes for beta-giardin and median body protein are (i) present as single copies in the genome, (ii) transcribed with very short 5' leader sequences. Two search algorithms designed to extract conserved motifs from either aligned or non-aligned sequences independently discovered three sites constituting a common pattern in all seven promoters. Sites were optimally aligned using weight matrix building trials to achieve the maximum 'information content'. Profiling the information content of best alignments defines the extent of the homologies as: a 9 bp box (initiator) at the start site and upstream 18 and 6 bp boxes. The initiator is the most highly conserved element and contains a universal Py-A-Pu motif at which transcription starts. We show that the best matrices can be combined in a search pattern that correctly locates transcription start sites in genomic DNA sequences.
机译:古代真核生物贾第鞭毛虫中的蛋白质编码基因缺乏典型的启动子共有元件。我们已经分析了七个具有相关功能的基因(结构性细胞骨架蛋白)的立即5'侧翼序列,以识别可能在转录起始中起作用的共享DNA图案。先前已经确定了五个基因的转录起始位点。基因组作图和mRNA引物延伸实验另外证明,β-贾第蛋白和中位体蛋白的基因(i)以单拷贝形式存在于基因组中,(ii)用非常短的5'前导序列转录。设计用于从比对或不比对序列中提取保守基序的两种搜索算法独立地发现了在所有七个启动子中构成共同模式的三个位点。使用权重矩阵构建试验对站点进行最佳对齐,以实现最大的“信息量”。对最佳比对的信息内容进行概要分析,将同源性的程度定义为:起始位点有9 bp的盒子(启动子),上游有18 bp和6 bp的盒子。引发剂是保守性最高的元素,并包含一个普遍的Py-A-Pu基序,从该基序开始转录。我们显示最好的矩阵可以组合在正确定位基因组DNA序列中转录起始位点的搜索模式中。

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