Park Seunghye, Lee Yew, Lee Jae-Hyeok, Jin EonSeon
Department of Life Science, Hanyang University, Seoul, South Korea.
Department of Botany, University of British Columbia, Vancouver, Canada.
Planta. 2013 Dec;238(6):1147-56. doi: 10.1007/s00425-013-1955-4. Epub 2013 Sep 17.
The development of highly inducible promoters is critical for designing effective transformation systems for transgenic analyses. In this study, we investigated the promoter of the light-inducible protein gene (LIP) of the marine alga Dunaliella sp. LIPs are homologs of the early light-induced proteins (ELIPs) of Arabidopsis thaliana. DNA sequence analysis revealed that the LIP promoter contains several light-responsive motifs. Constructs containing progressive truncations of the LIP promoter fused with a Renilla luciferase gene were introduced into Chlamydomonas reinhardtii to identify the light-responsive region in the promoter. Transcription from the LIP promoter was stimulated by high light (HL) in a light intensity-dependent manner. In contrast, oxidative stress induced by chemicals had little effect on the LIP promoter, which implies that the LIP promoter is exclusively induced by high light. Truncation of the promoter to a -100 base pair (bp) region abrogated light inducibility, which suggests the presence of a negative cis-regulatory element upstream of the -100 bp fragment. The LIP promoter can be utilized in transgenic research to specifically select and propagate transgenic microalgae under high-light conditions.
高度可诱导启动子的开发对于设计用于转基因分析的有效转化系统至关重要。在本研究中,我们研究了海洋藻类杜氏藻(Dunaliella sp.)的光诱导蛋白基因(LIP)的启动子。LIPs是拟南芥早期光诱导蛋白(ELIPs)的同源物。DNA序列分析表明,LIP启动子包含几个光响应基序。将含有与海肾荧光素酶基因融合的LIP启动子逐步截短的构建体导入莱茵衣藻(Chlamydomonas reinhardtii),以鉴定启动子中的光响应区域。LIP启动子的转录受到高光(HL)以光强度依赖的方式刺激。相比之下,化学物质诱导的氧化应激对LIP启动子影响很小,这意味着LIP启动子仅由高光诱导。将启动子截短至-100碱基对(bp)区域消除了光诱导性,这表明在-100 bp片段上游存在负性顺式调控元件。LIP启动子可用于转基因研究,以在高光条件下特异性选择和繁殖转基因微藻。