Protocols and programs for high-throughput growth and aging phenotyping in yeast.
Protocols and programs for high-throughput growth and aging phenotyping in yeast.
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DOI：
10.1371/journal.pone.0119807
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Linster CL
中科院分区：
文献类型：
--
作者：
Jung PP;Christian N;Kay DP;Skupin A;Linster CL
在微生物中，特别是在酵母中，标准表型分析方法包括通过在不同条件下测定生长速率来分析适合度。一种将高通量与高分辨率相结合的生长测定涉及在恒温和摇床读数器中从 96 孔板微培养物生成生长曲线。为了将该方法的通量提高到一个新的水平，我们在本研究中对其进行了调整，以使用 384 孔板。与 96 孔板或分批培养物相比，在 384 孔板中进行的实验之间提取的生长参数值（滞后时间、倍增时间和生物量产量）相关性良好，验证了表型筛选的更高通量方法。该方法不限于使用芽殖酵母酿酒酵母，如选自半子囊菌纲的其他物种的一致结果所示。此外，我们使用 384 孔板微培养来开发和验证酵母时间寿命 (CLS) 的更高通量测定，该参数仍然通常通过基于“菌落形成单位”计数的繁琐方法来确定。为了加速对所述生长和老化测定生成的大型数据集的分析，我们开发了免费的软件工具 GATHODE 和 CATHODE。这些工具允许从典型的读板机输出文件中半自动确定生长参数和 CLS 行为。所描述的方案和程序将提高许多基于酵母的系统遗传学实验以及各种类型的筛选的时间和成本效率。
In microorganisms, and more particularly in yeasts, a standard phenotyping approach consists in the analysis of fitness by growth rate determination in different conditions. One growth assay that combines high throughput with high resolution involves the generation of growth curves from 96-well plate microcultivations in thermostated and shaking plate readers. To push the throughput of this method to the next level, we have adapted it in this study to the use of 384-well plates. The values of the extracted growth parameters (lag time, doubling time and yield of biomass) correlated well between experiments carried out in 384-well plates as compared to 96-well plates or batch cultures, validating the higher-throughput approach for phenotypic screens. The method is not restricted to the use of the budding yeast Saccharomyces cerevisiae, as shown by consistent results for other species selected from the Hemiascomycete class. Furthermore, we used the 384-well plate microcultivations to develop and validate a higher-throughput assay for yeast Chronological Life Span (CLS), a parameter that is still commonly determined by a cumbersome method based on counting “Colony Forming Units”. To accelerate analysis of the large datasets generated by the described growth and aging assays, we developed the freely available software tools GATHODE and CATHODE. These tools allow for semi-automatic determination of growth parameters and CLS behavior from typical plate reader output files. The described protocols and programs will increase the time- and cost-efficiency of a number of yeast-based systems genetics experiments as well as various types of screens.
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影响因子：
29
作者：
Longo VD;Shadel GS;Kaeberlein M;Kennedy B
通讯作者：
Kennedy B
DOI：
10.1534/g3.111.000745
发表时间：
2012-02
期刊：
G3 (Bethesda, Md.)
影响因子：
--
作者：
Louis VL;Despons L;Friedrich A;Martin T;Durrens P;Casarégola S;Neuvéglise C;Fairhead C;Marck C;Cruz JA;Straub ML;Kugler V;Sacerdot C;Uzunov Z;Thierry A;Weiss S;Bleykasten C;De Montigny J;Jacques N;Jung P;Lemaire M;Mallet S;Morel G;Richard GF;Sarkar A;Savel G;Schacherer J;Seret ML;Talla E;Samson G;Jubin C;Poulain J;Vacherie B;Barbe V;Pelletier E;Sherman DJ;Westhof E;Weissenbach J;Baret PV;Wincker P;Gaillardin C;Dujon B;Souciet JL
通讯作者：
Souciet JL
芽殖酵母酿酒酵母乙醇耐受性的遗传解析
影响因子：
3.3
作者：
Hu, X. H.;Wang, M. H.;Luo, Z. W.
通讯作者：
Luo, Z. W.
影响因子：
64.8
作者：
MORTIMER, RK;JOHNSTON, JR
通讯作者：
JOHNSTON, JR
影响因子：
10.5
作者：
Kaeberlein, M;McVey, M;Guarente, L
通讯作者：
Guarente, L