STRENGTH AND TISSUE-SPECIFICITY OF CHIMERIC PROMOTERS DERIVED FROM THE OCTOPINE AND MANNOPINE SYNTHASE GENES
STRENGTH AND TISSUE-SPECIFICITY OF CHIMERIC PROMOTERS DERIVED FROM THE OCTOPINE AND MANNOPINE SYNTHASE GENES
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DOI：
10.1046/j.1365-313x.1995.7040661.x
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发表时间：
1995-04-01
期刊：
影响因子：
7.2
通讯作者：
GELVIN, SB
中科院分区：
文献类型：
--
作者：
NI, M;CUI, D;GELVIN, SB
为了研究启动子强度和组织特异性表达模式，将含有甘露糖醇合成酶（mas2’）亚域和八爪氨酸合成酶（ocs）启动子和激活子区域的嵌合启动子附着在β -葡萄糖醛酸酶报告基因上，并将其构建引入烟草植株。将ocs上游激活序列（UAS）的三聚体添加到mas启动子/激活子区域后，在所检测的所有组织中GUS活性水平均显著升高。在叶片组织中，该嵌合启动子的强度分别是CaMV 35S和增强双CaMV 35S启动子的156倍和26倍。由mas和ocs启动子/激活子引导的GUS活性的表达仅限于特定的细胞类型。将ocs或mas UAS添加到mas或ocs启动子/激活子区域可调节这些表达模式。将ocs UAS的三聚体添加到mas启动子/激活子区域，产生了一个转录控制元件，该元件在大多数细胞类型中指导GUS表达。除了在转基因烟草植株中的强表达外，该启动子在瞬时转化烟草叶盘和悬浮培养细胞以及木薯和豇豆外植体中的GUS表达水平高于CaMV 35S启动子。我们提出，含有三聚体的ocs UAS与mas启动子/激活子相结合的强启动子将有助于各种植物物种中连锁基因的高水平组成性表达。
To investigate promoter strength and the tissue-specific patterns of expression, chimeric promoters incorporating subdomains of mannopine synthase (mas2') and octopine synthase (ocs) promoter and activator regions were affixed to a beta-glucuronidase reporter gene and the constructions introduced into tobacco plants. Addition of a trimer of the ocs upstream activating sequence (UAS) to a mas promoter/activator region resulted in highly elevated levels of GUS activity in all tissues examined. In leaf tissue, this chimeric promoter is approximately 156-fold and 26-fold stronger than are the CaMV 35S and the 'enhanced' double CaMV 35S promoters, respectively. Expression of GUS activity directed by the mas and ocs promoters/activators is limited to specific cell types. Addition of the ocs or mas UAS to the mas or ocs promoter/activator regions modulated these expression patterns. The addition of a trimer of the ocs UAS to the mas promoter/activator region resulted in a transcriptional control element that directed GUS expression in most cell types. In addition to the strong expression in transgenic tobacco plants, this novel promoter directed higher levels of GUS expression than did the CaMV 35S promoter in transiently transformed tobacco leaf discs and suspension culture cells, as well as in cassava and cowpea explants. It is proposed that the strong promoter containing a trimer of the ocs UAS affixed to the mas promoter/activator will be useful for the very high level constitutive expression of linked genes in a wide variety of plant species.