Gudynaite-Savitch Loreta, Johnson Douglas A, Miki Brian L A
Ottawa-Carleton Institute of Biology, University of Ottawa, Ottawa, ON, Canada.
Plant Biotechnol J. 2009 Jun;7(5):472-85. doi: 10.1111/j.1467-7652.2009.00416.x.
The expression pattern of tissue-specific promoters in transgenes can be influenced by promoter/enhancer elements employed for the expression of selectable marker genes or elements found in DNA flanking the insertion site. We have developed an analytical system in Arabidopsis thaliana to investigate strategies useful in blocking or reducing nonspecific interactions. These experiments confirm that the DNA configuration and the insertion of spacer DNA aid in the appropriate expression of tissue-specific promoters. It is also demonstrated that the novel tobacco cryptic promoter (tCUP), when used to replace the cauliflower mosaic virus (CaMV) 35S promoter/enhancer, does not show nonspecific interactions. Furthermore, it is shown that insulators isolated from yeast and animals may have potential application in plants. Our results may allow the design of strategies that, individually or in combination, can be used to minimize nonspecific interactions and to design vectors for individual tissue-specific promoters.
转基因中组织特异性启动子的表达模式可能会受到用于表达选择标记基因的启动子/增强子元件或插入位点侧翼DNA中发现的元件的影响。我们在拟南芥中开发了一种分析系统，以研究有助于阻断或减少非特异性相互作用的策略。这些实验证实，DNA构型和间隔DNA的插入有助于组织特异性启动子的适当表达。还证明，当用于取代花椰菜花叶病毒（CaMV）35S启动子/增强子时，新型烟草隐蔽启动子（tCUP）不会表现出非特异性相互作用。此外，研究表明，从酵母和动物中分离出的绝缘子可能在植物中具有潜在应用。我们的结果可能有助于设计单独或组合使用的策略，以尽量减少非特异性相互作用，并为单个组织特异性启动子设计载体。
