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首页> 外文期刊>Molecular and Cellular Biology >Selection of novel exon recognition elements from a pool of random sequences.
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Selection of novel exon recognition elements from a pool of random sequences.

机译：从随机序列库中选择新型外显子识别元件。

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A 20-nucleotide sequence close to the 3' end of the internal exon of a model two-intron, three-exon pre-mRNA (DUP184 [Z. Dominski and R. Kole, J. Biol. Chem. 269:23590-23596, 1994]) was replaced by a random 20-mer, resulting in a pool of pre-mRNAs which, like the initial DUP184 construct, were spliced in vitro by a pathway leading to predominant skipping of the internal exon. The randomized pre-mRNAs were subjected to a selection protocol, resulting in a pool enriched in pre-mRNAs that efficiently included the internal exon. Isolation and sequencing of a number of clones corresponding to the selected pre-mRNAs showed that two classes of sequences were selected from the initial pool. Most abundant among these were sequences with purine tracts similar to those in the recently identified exon-splicing enhancers while a smaller class included sequences lacking discernible purine tracts within the 20-nucleotide region. Splicing of selected pre-mRNAs showed that the purine tracts vary in their ability to promote exon inclusion and, more important, that sequences lacking purine tracts stimulate inclusion of the internal exon as efficiently as their purine-rich counterparts. The latter result indicates the existence of a novel class of exon recognition sequences or splicing enhancers.
机译：一个20核苷酸序列，靠近模型两个内含子，三个外显子的pre-mRNA内部外显子的3'端（DUP184 [Z. Dominski and R.Kole，J.Biol.Chem.269：23590-23596 ，1994]）替换为随机的20-mer，产生大量的pre-mRNA，与最初的DUP184构建体一样，通过导致内部外显子显着跳过的途径在体外进行剪接。对随机化的前mRNA进行选择方案，产生一个富含前mRNA的库，该库有效地包含内部外显子。对应于所选前mRNA的许多克隆的分离和测序表明，从初始库中选择了两类序列。其中最丰富的是具有与最近鉴定的外显子剪接增强子相似的嘌呤束的序列，而较小的一类包含在20个核苷酸区域内缺乏可辨别的嘌呤束的序列。选定的前mRNA的剪接表明，嘌呤束促进外显子包涵的能力各不相同，更重要的是，缺乏嘌呤束的序列与富含嘌呤的对应物一样有效地刺激内部外显子的包涵。后一结果表明存在一类新的外显子识别序列或剪接增强子。

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