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编码类囊体蛋白基因5'-非翻译前导序列的片段含有转录所必需的顺式元件。

Segments encoding 5'-untranslated leaders of genes for thylakoid proteins contain cis-elements essential for transcription.

作者信息

Bolle C, Sopory S, Lübberstedt T, Herrmann R G, Oelmüller R

机构信息

Botanisches Institut, Ludwig-Maximilians-Universität, München, Germany.

出版信息

Plant J. 1994 Oct;6(4):513-23. doi: 10.1046/j.1365-313x.1994.6040513.x.

Abstract

The promoter region -118/-29 of the spinach PetH gene encoding the ferredoxin-NADP(+)-oxidoreductase contains crucial cis-elements for the regulated expression, while sequences for the 5'-untranslated leader determine the quantitative expression of chimeric GUS gene fusions in transgenic tobacco. Deletion of leader sequences in chimeric GUS gene fusions of the spinach PetE and PsaF genes (for plastocyanin and the subunit III of photosystem I, respectively) results also in a decline in the GUS activity. Appropriate gene constructs and run-on transcription assays demonstrate unambiguously that the leaders of all three genes are involved in transcription rather than in post-transcriptional processes. They appear to contain gene-specific control elements rather than cis-determinants for general initiation factors. Expression-relevant segments in the PsaF and PetH leaders contain two CT-rich sequences, designated CT-LB and CT-B, of which at least the former binds to a protein factor in gel mobility shift assays. These motifs are not found in the PetE leader. The findings imply that leader sequences may contain cis-elements that are essential for the transcription, that they influence GUS gene expression quantitatively rather than qualitatively, and that these elements, as those of promoters, can be quite variable in sequence.

摘要

编码铁氧还蛋白-NADP(+)-氧化还原酶的菠菜PetH基因的启动子区域-118/-29含有调控表达的关键顺式元件,而5'-非翻译前导序列决定了转基因烟草中嵌合GUS基因融合体的定量表达。菠菜PetE和PsaF基因(分别用于质体蓝素和光系统I的亚基III)的嵌合GUS基因融合体中前导序列的缺失也导致GUS活性下降。合适的基因构建体和连续转录分析明确表明,这三个基因的前导序列均参与转录而非转录后过程。它们似乎含有基因特异性控制元件,而非一般起始因子的顺式决定簇。PsaF和PetH前导序列中与表达相关的片段含有两个富含CT的序列,分别命名为CT-LB和CT-B,其中至少前者在凝胶迁移率变动分析中能与一种蛋白质因子结合。这些基序在PetE前导序列中未发现。这些发现表明,前导序列可能含有对转录至关重要的顺式元件,它们对GUS基因表达的影响是定量而非定性的,并且这些元件与启动子元件一样,在序列上可能有很大差异。

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