Rouster J, van Mechelen J, Cameron-Mills V
Carlsberg Research Laboratory, Copenhagen, Denmark.
Plant J. 1998 Aug;15(3):435-40. doi: 10.1046/j.1365-313x.1998.00193.x.
The barley lipoxygenase 1 (Lox1) gene encodes a protein expressed in embryos during grain development and germination and in leaves after methyl-jasmonate (MeJA) treatment. Transient gene expression assays in germinating barley embryos were used to identify cis-regulatory elements involved in the embryo-specific expression of the Lox1 gene. Analysis of transcriptional or translational fusions between Lox1 5' upstream sequences and the gusA reporter gene indicated that the 5'-untranslated leader sequence was involved in embryo-specific expression. Replacement of the leader sequence from the aleurone-specific Chi26 gene with the Lox1 leader sequence resulted in a chimeric gene expressed at high levels in embryo as well as in aleurone cells. Insertion of the Lox1 leader sequence between the 35S minimum promoter (A domain -90/+8) and the gusA reporter gene greatly enhanced promoter activity in a tissue-specific manner. Deletion/replacement analysis of the Lox1 leader sequence, combined with transient expression in germinating embryos and in vitro transcription/translation assays, suggests that the Lox1 leader sequence contains cis-elements regulating qualitative (tissue-specific) and quantitative gene expression.
大麦脂氧合酶1(Lox1)基因编码一种在籽粒发育和萌发过程中的胚以及茉莉酸甲酯(MeJA)处理后的叶片中表达的蛋白质。利用发芽大麦胚中的瞬时基因表达分析来鉴定参与Lox1基因胚特异性表达的顺式调控元件。对Lox1 5'上游序列与gusA报告基因之间的转录或翻译融合分析表明,5'-非翻译前导序列参与胚特异性表达。用Lox1前导序列替换糊粉层特异性Chi26基因的前导序列,产生了一个在胚以及糊粉层细胞中高水平表达的嵌合基因。在35S最小启动子(A结构域-90/+8)和gusA报告基因之间插入Lox1前导序列以组织特异性方式极大地增强了启动子活性。对Lox1前导序列的缺失/替换分析,结合在发芽胚中的瞬时表达和体外转录/翻译分析,表明Lox1前导序列含有调控定性(组织特异性)和定量基因表达的顺式元件。