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豌豆凝集素启动子中的22bp W1元件对于烟草种子中的高基因表达是必需的,并且作为多聚体时是充分的。

The 22 bp W1 element in the pea lectin promoter is necessary and, as a multimer, sufficient for high gene expression in tobacco seeds.

作者信息

de Pater S, Pham K, Klitsie I, Kijne J

机构信息

Center for Phytotechnology, Leiden University, AL Leiden, Netherlands.

出版信息

Plant Mol Biol. 1996 Nov;32(3):515-23. doi: 10.1007/BF00019103.

Abstract

The pea lectin (Psl) gene encodes an abundant seed protein. Its seed-specific expression pattern is conserved in transgenic tobacco plants. Progressive 5' promoter deletions resulted in a gradual decrease of transcriptional activity in tobacco seed. A fragment of 115 bp still conferred seed-specific expression albeit at a low level. This fragment contains a 22 bp element (W1), which has been demonstrated to be important for seed-specific expression when coupled as a trimer to a heterologous TATA box (de Pater et al., Plant Cell 5:877-886, 1993). Here we show that deletion of W1 in the natural promoter context resulted in a strongly decreased level of gene expression. A 4 bp mutation of W1 reduced the expression of truncated derivatives of the Psl promoter. A single copy of W1 coupled to the TATA box of the CaMV 35S promoter directed low gene expression in seeds and leaves. Multimerization enhanced the expression in seeds up to 100-fold, to levels found with the Psl promoter, whereas the expression level in leaves remained low. These results demonstrate that the W1 element is an essential control element in the Psl promoter. When taken out of its natural context and multimerized, it is sufficient for high expression in seeds.

摘要

豌豆凝集素(Psl)基因编码一种丰富的种子蛋白。其种子特异性表达模式在转基因烟草植株中得以保留。5' 端启动子的逐步缺失导致烟草种子中转录活性逐渐降低。一个115 bp的片段仍能赋予种子特异性表达,尽管表达水平较低。该片段包含一个22 bp的元件(W1),当它作为三聚体与异源TATA框偶联时,已被证明对种子特异性表达很重要(德·帕特等人,《植物细胞》5:877 - 886,1993)。在此我们表明,在天然启动子背景下缺失W1会导致基因表达水平大幅下降。W1的一个4 bp突变降低了Psl启动子截短衍生物的表达。单个拷贝的W1与CaMV 35S启动子的TATA框偶联,在种子和叶片中导致低水平的基因表达。多聚化使种子中的表达增强至100倍,达到Psl启动子的表达水平,而叶片中的表达水平仍然较低。这些结果表明,W1元件是Psl启动子中的一个关键调控元件。当将其从天然背景中取出并多聚化时,它足以在种子中实现高表达。

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