Phenomic assessment of genetic buffering by kinetic analysis of cell arrays.
Phenomic assessment of genetic buffering by kinetic analysis of cell arrays.
复制标题
通过细胞阵列动力学分析对遗传缓冲进行表型评估。
DOI：
10.1007/978-1-4939-1363-3_12
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发表时间：
2014
期刊：
影响因子：
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通讯作者：
Hartman4th,JohnL
中科院分区：
文献类型：
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作者：
Rodgers,John;Guo,Jingyu;Hartman4th,JohnL
定量高通量细胞阵列表型分析（Q-HTCP）应用于酵母基因缺失突变体的基因组收集，用于系统、全面地评估基因和基因组合对任何感兴趣的表型的贡献（表型分析）。相互作用的基因网络影响每一种表型。遗传缓冲是指基因相互作用网络如何稳定或破坏表型。与基因组学一样，表型组学的分辨率也各不相同，因为在每个样本分配更多的测量次数以增强表型的量化与通过每个样本获得更少的测量次数来增加不同样本的数量之间存在权衡。我们描述的Q-HTCP方案通过从琼脂细胞阵列的时间序列成像中获得动态生长曲线来评估每个实验的50,000-70,000个培养物。这种方法是为酵母基因缺失菌株开发的，但它也可以应用于在固体琼脂培养基上生长的其他微生物突变体阵列。我们描述的方法包括冷冻原液的创建和维护、液源阵列的制备、琼脂靶板的印刷、图像扫描、图像分析、曲线拟合和基因相互作用的评估。
Quantitative high-throughput cell array phenotyping (Q-HTCP) is applied to the genomic collection of yeast gene deletion mutants for systematic, comprehensive assessment of the contribution of genes and gene combinations to any phenotype of interest (phenomic analysis). Interacting gene networks influence every phenotype. Genetic buffering refers to how gene interaction networks stabilize or destabilize a phenotype. Like genomics, phenomics varies in its resolution with there being a trade-off allocating a greater number of measurements per sample to enhance quantification of the phenotype vs. increasing the number of different samples by obtaining fewer measurements per sample. The Q-HTCP protocol we describe assesses 50,000–70,000 cultures per experiment by obtaining kinetic growth curves from time series imaging of agar cell arrays. This approach was developed for the yeast gene deletion strains, but it could be applied as well to other microbial mutant arrays grown on solid agar media. The methods we describe are for creation and maintenance of frozen stocks, liquid source array preparation, agar destination plate printing, image scanning, image analysis, curve fitting, and evaluation of gene interaction.