National Institute for Basic Biology, Okazaki, Japan ; ERATO, Japan Science and Technology Agency, Okazaki, Japan ; Plant Biotechnology, Faculty of Biology, University of Freiburg, Freiburg, Germany ; FRIAS - Freiburg Institute for Advanced Studies, Freiburg, Germany.
PLoS One. 2013 Sep 27;8(9):e77356. doi: 10.1371/journal.pone.0077356. eCollection 2013.
Inducible transgene expression provides a useful tool to analyze gene function. The moss Physcomitrellapatens is a model basal land plant with well-developed research tools, including a high efficiency of gene targeting and substantial genomics resources. However, current systems for controlled transgene expression remain limited. Here we report the development of an estrogen receptor mediated inducible gene expression system, based on the system used in flowering plants. After identifying the appropriate promoters to drive the chimeric transducer, we succeeded in inducing transcription over 1,000-fold after 24 h incubation with β-estradiol. The P. patens system was also effective for high-level long-term induction of gene expression; transcript levels of the activated gene were maintained for at least seven days on medium containing β-estradiol. We also established two potentially neutral targeting sites and a set of vectors for reproducible expression of two transgenes. This β-estradiol-dependent system will be useful to test genes individually or in combination, allowing stable, inducible transgenic expression in P. patens.
诱导型转基因表达为分析基因功能提供了一种有用的工具。藓苔Physcomitrellapatens 是一种具有完善研究工具的基础陆地植物模式生物，包括高效的基因靶向和大量的基因组资源。然而，目前控制转基因表达的系统仍然有限。在这里，我们报告了一种基于开花植物中使用的系统的雌激素受体介导的诱导型基因表达系统的开发。在确定了合适的启动子来驱动嵌合转导子之后，我们成功地在 24 小时β-雌二醇孵育后诱导转录超过 1000 倍。P. patens 系统也可有效进行高水平的长期基因表达诱导；在含有β-雌二醇的培养基中，激活基因的转录本水平至少维持七天。我们还建立了两个潜在的中性靶向位点和一套用于两个转基因可重复表达的载体。这种依赖于β-雌二醇的系统将有助于单独或组合测试基因，允许在 P. patens 中稳定、诱导型的转基因表达。
