摘要
建立了应用流式细胞仪分选植物特定类型细胞的方法。以拟南芥(Arabidopsis thaliana)Wer::GFP转基因株系为材料,用激光共聚焦显微镜鉴定GFP的表达位置,采用酶解法制备拟南芥根尖原生质体,应用流式细胞仪荧光激活细胞分选技术(FACS)分选收集GFP阳性细胞,并提取细胞的RNA。结果表明,Wer::GFP转基因株系仅在根表皮发育早期的非根毛细胞中表达GFP;利用酶解法制备的根尖原生质体数目较多;从FACS分选收集的细胞中提取的RNA质量较好,可用于研究特定类型细胞的基因表达谱。应用流式细胞仪分选拟南芥非根毛细胞的方法为研究植物特定类型细胞的基因表达谱及基因功能奠定了技术基础。
We developed a method for sorting specific cells from Arabidopsis thaliana.We used Wer：：GFP transgenic lines as the experimental material;laser scanning confocal microscopy was used to identify the green fluorescent protein（GFP） location;enzymatic digestion was used to prepare root tip protoplasts;fluorescence-activated cell sorting（FACS） was used to collect GFP-positive cells;and total RNA was extracted from these cells.Only the early developmental non-root hair cells in the root epidermis expressing GFP in Wer：：GFP transgenic lines.A large number of root-tip protoplasts were obtained by enzymatic digestion,and GFP-positive cells were successfully collected by FACS.The RNA extracted from these specific cells could be used for gene expression profiling.We have established a successful method for sorting specific cells by flow cytometry.
出处
《植物学报》
CAS
CSCD
北大核心
2010年第4期460-465,共6页
Chinese Bulletin of Botany
基金
国家自然科学基金(No.30771094)
北京市自然科学基金(No.5082003)
关键词
拟南芥
流式细胞仪
荧光激活细胞分选
非根毛细胞
原生质体
Arabidopsis thaliana
flow cytometry
fluorescent-activated cell sorting
non-hair cell
protoplast
