Department of Plant Protection, Shenyang Agriculture University, Shenyang 110161, China ; Guangxi Crop Genetic Improvement and Biotechnology Key Lab, Nanning 530007, China.
Department of Plant Protection, Shenyang Agriculture University, Shenyang 110161, China.
Saudi J Biol Sci. 2014 Apr;21(2):185-90. doi: 10.1016/j.sjbs.2013.09.011. Epub 2013 Oct 9.
In this study, optimized 2-DE sample preparation methodologies were established for suspension-cultured ginseng cells. Three commonly used protein extraction methods (Trichloroacetic acid-acetone, urea/thiourea and phenol extraction method) were evaluated for proteomic analysis of suspension cultures of ginseng. A comparative analysis of suspension-cultured ginseng cells proteome induced by salicylic acid (SA) was reported. The results demonstrated that phenol extraction method was the best method based on protein extraction efficiency and the good quality of 2-DE patterns for suspension-cultured ginseng cells. Fifteen differentially expressed proteins induced by salicylic acid in suspension-cultured ginseng cells were identified by MALDI-TOF-MS. These identified proteins were involved in defense and stress response, energy metabolism, signal transduction/transcription, protein synthesis and metabolism, and photosynthesis. Chaperonin 60, related to defense responses, was more abundant in suspension-cultured ginseng cells after application of SA. Vacuolar ATPase subunit B was newly induced in SA treatment.
在这项研究中，建立了优化的人参悬浮细胞 2-DE 样品制备方法。为了进行人参悬浮培养的蛋白质组学分析，评估了三种常用的蛋白质提取方法（三氯乙酸-丙酮法、尿素/硫脲法和酚提取法）。报道了水杨酸（SA）诱导的人参悬浮细胞蛋白质组的比较分析。结果表明，基于蛋白质提取效率和人参悬浮细胞 2-DE 图谱的质量，酚提取法是最佳方法。通过 MALDI-TOF-MS 鉴定了 15 种由水杨酸诱导的人参悬浮细胞差异表达蛋白。这些鉴定的蛋白参与防御和应激反应、能量代谢、信号转导/转录、蛋白质合成和代谢以及光合作用。与防御反应有关的热休克蛋白 60 在应用 SA 后在人参悬浮细胞中更为丰富。液泡 ATP 酶亚基 B 在 SA 处理时新诱导。
