Kirke Justin, Kaplan Noah, Velez Stephanie, Jin Xiao-Lu, Vichyavichien Paveena, Zhang Xing-Hai
Department of Biological Sciences, Florida Atlantic University, Boca Raton, FL, 33431, USA.
Mol Biotechnol. 2018 Mar;60(3):194-202. doi: 10.1007/s12033-018-0060-0.
A promoter is an essential structural component of a gene that controls its transcription activity in different development stages and in response to various environmental stimuli. Knowledge of promoter functionality in heterologous systems is important in the study of gene regulation and biotechnological application. In order to explore the activity of the pepper capsaicin synthase gene (PUN1) promoter, gene constructs of pPUN1::GUS (for β-glucuronidase) and pPUN1::NtKED (for a tobacco wound-responsive protein) were introduced into tobacco and tomato, respectively, and their activities were examined. Higher levels of GUS staining intensity and transcription were detected in ovary, anther and pollen than other tissues or organs in tobacco plants. Likewise, transgenic tomato fruits had a higher level of pPUN1::NtKED gene expression than the leaf and flower. The PUN1-driven gene expression can be transiently induced by wounding, heat (40 °C) and the capsaicinoid biosynthetic pathway precursor phenylalanine. When compared to the reported pPUN1::GUS-expressing Arabidopsis, the PUN1 promoter exhibited a more similar pattern of activities among pepper, tobacco and tomato, all Solanaceae plants. Our results suggest the potential utility of this tissue-preferential and inducible promoter in other non-pungent Solanaceae plants for research of gene function and regulation as well as in the biotechnological applications.
启动子是基因的一个重要结构组成部分,它在不同发育阶段以及对各种环境刺激的响应中控制基因的转录活性。了解异源系统中启动子的功能对于基因调控研究和生物技术应用很重要。为了探究辣椒辣椒素合成酶基因(PUN1)启动子的活性,分别将pPUN1::GUS(用于β-葡萄糖醛酸酶)和pPUN1::NtKED(用于烟草伤口响应蛋白)的基因构建体导入烟草和番茄中,并检测它们的活性。在烟草植株的子房、花药和花粉中检测到的GUS染色强度和转录水平高于其他组织或器官。同样,转基因番茄果实中pPUN1::NtKED基因的表达水平高于叶片和花朵。PUN1驱动的基因表达可被伤口、热(40°C)和辣椒素生物合成途径前体苯丙氨酸短暂诱导。与已报道的表达pPUN1::GUS的拟南芥相比,PUN1启动子在辣椒、烟草和番茄(均为茄科植物)中表现出更相似的活性模式。我们的结果表明,这种组织特异性和可诱导的启动子在其他非辛辣茄科植物中对于基因功能和调控研究以及生物技术应用具有潜在用途。