Department of Plant Science, College of Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea; Department of Horticultural Science, Kyungpook National University, Daegu, Republic of Korea.
Biotechnology Institute, Nongwoo Bio Co, Ltd, Yeoju, Republic of Korea; Department of Plant Biology, Cornell University, Ithaca, NY, USA.
Biochem Biophys Res Commun. 2014 Apr 18;446(4):882-8. doi: 10.1016/j.bbrc.2014.03.022. Epub 2014 Mar 13.
Plant cells secrete diverse sets of constitutively- and conditionally-expressed proteins under various environmental and developmental states. Secreted protein populations, or secretomes have multiple functions, including defense responses, signaling, metabolic processes, and developmental regulation. To identify genes encoding secreted proteins that function in fruit development and ripening, a yeast secretion trap (YST) screen was employed using pepper (Capsicum annuum) fruit cDNAs. The YST screen revealed 80 pepper fruit-related genes (CaPFRs) encoding secreted proteins including cell wall proteins, several of which have not been previously described. Transient GFP-fusion assay and an in planta secretion trap were used to validate the secretion of proteins encoded by selected YST clones. In addition, RNA gel blot analyses provided further insights into their expression and regulation during fruit development and ripening. Integrating our data, we conclude that the YST provides a valuable functional genomics tool for the identification of substantial numbers of novel secreted plant proteins that are associated with biological processes, including fruit development and ripening.
在各种环境和发育状态下，植物细胞会分泌多种组成型和条件型表达的蛋白质。分泌蛋白群体（或称为分泌组）具有多种功能，包括防御反应、信号转导、代谢过程和发育调控。为了鉴定在果实发育和成熟过程中起作用的编码分泌蛋白的基因，我们使用辣椒（Capsicum annuum）果实 cDNA 进行了酵母分泌陷阱（YST）筛选。YST 筛选揭示了 80 个与辣椒果实相关的基因（CaPFRs），这些基因编码分泌蛋白，包括细胞壁蛋白，其中一些以前没有描述过。瞬时 GFP 融合分析和体内分泌陷阱被用来验证选定 YST 克隆编码的蛋白质的分泌。此外，RNA 凝胶印迹分析提供了进一步的见解，了解它们在果实发育和成熟过程中的表达和调控。综合我们的数据，我们得出结论，YST 为鉴定与生物过程（包括果实发育和成熟）相关的大量新的分泌植物蛋白提供了一种有价值的功能基因组学工具。
