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Isolation and characterization of a harvest-induced promoter of an alfalfa gene, hi7

机译:苜蓿基因hi7的收获诱导启动子的分离和鉴定

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摘要

A cDNA library was made by subtractive hybridization of cDNA from non-harvested and harvested alfalfa plants. One cDNA clone, hi7, in Northern blot analysis showed a high level of transcription of this gene in harvested tissue, but not in non-harvested tissue, nor in wounded or heat shock treated tissue. A 640 bp promoter region isolated by genomic walking based on hi7 cDNA sequence, was fused to a GUS gene and transferred to tobacco and Medicago truncatula. When leaf tissue from transgenic tobacco plants was harvested, incubated for 24 h at room temperature, and then stained for GUS activity, a strong blue colour was observed throughout the tissue, but not in tissue that was stained immediately following harvesting. The expression of the GUS gene was also induced in tobacco plants sprayed with abscissic acid (ABA) and in callus derived from these plants when transferred to medium containing ABA.
机译:通过对未收获和收获的苜蓿植物中的cDNA进行减性杂交来制作cDNA文库。在Northern印迹分析中,一个cDNA克隆hi7在收获的组织中显示了该基因的高水平转录,但在未收获的组织,受伤或受热激处理的组织中却没有。通过基于hi7 cDNA序列的基因组步移分离的640 bp启动子区域与GUS基因融合,并转移至烟草和截叶苜蓿。当收获转基因烟草植物的叶片组织,在室温下孵育24小时,然后对GUS活性染色时,在整个组织中均观察到了强烈的蓝色,但在收获后立即染色的组织中未观察到强烈的蓝色。当用脱落酸(ABA)喷洒的烟草植物以及从这些植物衍生的愈伤组织转移到含有ABA的培养基中时,也诱导了GUS基因的表达。

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