Large-scale Phenotypic Profiling of Gene Deletion Mutants in Candida glabrata
对光滑念珠菌中的基因缺失突变体进行大规模表型分析
(*contributed equally to this work) 发布: 2015年07月20日第5卷第14期 DOI: 10.21769/BioProtoc.1530 浏览次数: 10965
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参见作者原研究论文
The authors used this protocol in:
Jun 2014
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Abstract
Here, we describe a method enabling the phenotypic profiling of genome-scale deletion collections of fungal mutants to detect phenotypes for various stress conditions. These stress conditions include among many others antifungal drug susceptibility, temperature-induced and osmotic as well as heavy metal or oxidative stress. The protocol was extensively used to phenotype a collection of gene deletion mutants in the human fungal pathogen Candida glabrata (C. glabrata) (Schwarzmüller et al., 2014).
Keywords: Phenotyping (表型)
Antifungal Drug Susceptiility (抗真菌药物susceptiility)
Stress Phenotypes (压力表型)
Morphology (形态学)
Robotic Screening (机器人筛选)
Materials and Reagents
- C. glabrata strain (ATCC2001, clinical isolates)
- Sterile water (double distilled)
- Bacto™ peptone (BD Biosciences, catalog number: 211820 )
- Bacto™ yeast extract (BD Biosciences, catalog number: 212720 )
- Bacto™ agar (BD Biosciences, catalog number: 214030 )
- Difco™ yeast nitrogen base (YNB) (BD Biosciences, catalog number: 233520 )
- Glucose (Merck KGaA, catalog number: 108337 )
- Adenine (Sigma-Aldrich, catalog number: A8626 )
- L-arginine (Sigma-Aldrich, catalog number: A5006 )
- L-tyrosine (Sigma-Aldrich, catalog number: T3754 )
- L-isoleucine (Sigma-Aldrich, catalog number: I2752 )
- L-phenylalanine (Sigma-Aldrich, catalog number: P2126 )
- L-glutamic acid(Sigma-Aldrich, catalog number: G1251 )
- L-aspartic acid (Sigma-Aldrich, catalog number: A9256 )
- L-threonine (Sigma-Aldrich, catalog number: T8625 )
- L-serine (Sigma-Aldrich, catalog number: S4500 )
- L-valine (Sigma-Aldrich, catalog number: V0500 )
- L-methionine (Sigma-Aldrich, catalog number: M9625 )
- Glycerol (Sigma-Aldrich, catalog number: G5516 )
- YPD media (see Recipes)
- Solid YPD media (see Recipes)
- 2x YPD media (see Recipes)
- SC media (see Recipes)
- Solid SC media (see Recipes)
- Amino acid mix (see Recipes)
- 15% glycerol solution (see Recipes)
Equipment
- Square plates (PlusPlate) (40 ml/plate) (Singer Instruments, catalog number: PLU-001 )
- Pin Pad (RePad 96 Long) (Singer Instruments, catalog number: REP-001 )
- 96-well plates (200 µl/well) (Starlab, catalog number: CC76827596 )
- Petri dishes (92 mm) (SARSTEDT AG, catalog number: 82.1473 )
- Culture tubes (Starlab, catalog number: I14850810 )
- Yeast replica robot (Singer Instruments, catalog number: RoToR HDA )
- Plate reader (Victor3V) (PerkinElmer, catalog number: 1420040 )
- 48-well replica stamp (V&P Scientific, catalog number: VP 408H )
- 96-well replica stamp (V&P Scientific, catalog number: VP 407 )
- Library copier (guide for stamps) (V&P Scientific, model: VP 381 )
- Plate mixer (Eppendorf, catalog number: 5353000014 )
- Rotary shaker for culture flasks (innova44) (New Brunswick Scientific, catalog number: M1282-0002 )
- Incubator (Heraeus Instruments, catalog number: B6120 )
Procedure
文章信息
版权信息
© 2015 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Istel, F., Schwarzmüller, T., Tscherner, M. and Kuchler, K. (2015). Large-scale Phenotypic Profiling of Gene Deletion Mutants in Candida glabrata. Bio-protocol 5(14): e1530. DOI: 10.21769/BioProtoc.1530.
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分类
微生物学 > 抗微生物试验 > 抗真菌试验
系统生物学 > 基因组学 > 功能基因组学
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