Department of Life Science, Research Institute for Natural Sciences, Hanyang University, Seoul 04763, Republic of Korea.
Division of Polar Life Science, Korea Polar Research Institute, Korea Institute of Ocean Science and Technology, Incheon 21990, Republic of Korea.
J Microbiol Biotechnol. 2020 Nov 28;30(11):1777-1784. doi: 10.4014/jmb.2007.07024.
To increase the availability of microalgae as producers of valuable compounds, it is necessary to develop novel systems for gene expression regulation. Among the diverse expression systems available in microalgae, none are designed to induce expression by low temperature. In this study, we explored a cold-inducible system using the antifreeze protein (AFP) promoter from a polar diatom, . A vector containing the promoter () was generated to regulate nuclear gene expression, and reporter genes ( luciferase () and mVenus fluorescent protein ()) were successfully expressed in the model microalga, . In particular, under the control of , the expression of these genes was increased at low temperature, unlike , a promoter that is widely used for gene expression in . Promoter truncation assays showed that cold inducibility was still present even when was shortened to 600 bp, indicating the presence of a low-temperature response element between -600 and -477 bp. Our results show the availability of new heterologous gene expression systems with cold-inducible promoters and the possibility to find novel low-temperature response factors in microalgae. Through further improvement, this cold-inducible promoter could be used to develop more efficient expression tools.
为了提高微藻作为有价值化合物生产者的可用性，有必要开发新的基因表达调控系统。在微藻中可用的各种表达系统中，没有一个是专门设计用于低温诱导表达的。在这项研究中，我们探索了一种使用来自极地硅藻的抗冻蛋白 (AFP) 启动子的冷诱导系统。生成了一个包含 启动子 () 的载体来调节核基因表达，并且报告基因（荧光素酶 () 和 mVenus 荧光蛋白 ()）在模式微藻 中成功表达。特别是，在 的控制下，这些基因的表达在低温下增加，与广泛用于 的启动子不同。启动子截断分析表明，即使 缩短至 600 bp，冷诱导性仍然存在，表明在 -600 至 -477 bp 之间存在低温反应元件。我们的结果表明，具有冷诱导启动子的新型异源基因表达系统是可用的，并且有可能在微藻中找到新的低温反应因子。通过进一步改进，这个冷诱导启动子可以用于开发更有效的表达工具。
