Histostaining for Tissue Expression Pattern of Promoter-driven GUS Activity in Arabidopsis
用启动子融合GUS的方法分析组织水平上基因的表达模式
In Press 发布: 2011年07月05日 DOI: 10.21769/BioProtoc.93 浏览次数: 31378
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Abstract
Promoter-driven GUS (beta-glucuronidase) activity is the most commonly used technique for tissue-specific expression patterns in Arabidopsis. In this procedure, GUS enzyme converts 5-bromo-4-chloro-3-indolyl glucuronide (X-Gluc) to a blue product. The staining is very sensitive. Processed samples can be examined under dissecting microscope or Differential Interference Contrast (Nomaski) microscope for bright blue color over cleared transparent background. Note this assay does not provide accurate information to subcellular levels.
Keywords: Gene expression (基因表达)
GUS activity (GUS活性)
Histostaining (免疫组化)
Arabidopsis (拟南芥)
Promoter activity (启动子活性)
Materials and Reagents
- Transgenic plants that contain genomic integration of a promoter: GUS expression cassette
- Potassium Ferrocyanide
- Potassium Ferricyanide
- Triton X-100
- 50 mM NaHPO4 buffer (pH 7.2)
- Dimethylformamide (DMF)
- Acetone
- NaHPO4 buffer
- 5-bromo-4-chloro-3-indolyl beta-D-glucuronide cyclohexamine salt (X-Gluc)
- 200 proof ethanol (once opened, 200 proof becomes essentially 190 proof)
- Staining buffer (see Recipes)
- Stock solutions (see Recipes)
Equipment
- Eppendorf tubes
- Vacuum
- Dissecting or light microscope
- Differential Interference Contrast (Nomaski) microscope
Procedure
文章信息
版权信息
© 2011 The Authors; exclusive licensee Bio-protocol LLC.
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分类
细胞生物学 > 组织分析 > 组织染色
植物科学 > 植物细胞生物学 > 组织分析
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