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在指数生长的酵母细胞中存在一个稳定的转录组。

There is a steady-state transcriptome in exponentially growing yeast cells.

机构信息

Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Biológicas, Universitat de València, C/Dr. Moliner 50, 46100 Burjassot, Spain.

出版信息

Yeast. 2010 Jul;27(7):413-22. doi: 10.1002/yea.1768.

Abstract

The growth of yeast cells in batches in glucose-based media is a standard condition in most yeast laboratories. Most gene expression experiments are done by taking this condition as a reference. Presumably, cells are in a stable physiological condition that can be easily reproduced in other laboratories. With this assumption, however, it is necessary to consider that the average amount of the mRNAs per cell for most genes does not change during exponential growth. That is to say, there is a steady-state condition for the transcriptome. However, this has not been rigorously demonstrated to date. In this work we take several cell samples during the exponential phase growth to perform a kinetic study using the genomic run-on (GRO) technique, which allows simultaneous measurement of the amount of mRNA and transcription rate variation at the genomic level. We show here that the steady-state condition is fulfilled for almost all the genes during most exponential growth in yeast extract-peptone-dextrose medium (YPD) and, therefore, that simultaneous measures of the transcription rates and the amounts of mRNA can be used for indirect mRNA stability calculations. With this kinetic approach, we were also able to determine the relative influence of the transcription rate and the mRNA stability changes for the mRNA variation for those genes that deviate from the steady state.

摘要

在基于葡萄糖的培养基中分批培养酵母细胞是大多数酵母实验室的标准条件。大多数基因表达实验都是以这种条件作为参考进行的。推测细胞处于稳定的生理状态,在其他实验室中很容易重现。然而,根据这一假设,有必要考虑到大多数基因的每个细胞中的 mRNA 平均数量在指数生长期不会发生变化。也就是说,转录组存在一个稳态条件。然而,到目前为止,这还没有得到严格的证明。在这项工作中,我们在指数生长期取几个细胞样本,使用基因组运行-on(GRO)技术进行动力学研究,该技术允许在基因组水平上同时测量 mRNA 的数量和转录率变化。我们在这里表明,在酵母提取物-蛋白胨-葡萄糖培养基(YPD)中,大多数指数生长过程中,几乎所有基因都满足稳态条件,因此,可以同时测量转录率和 mRNA 量,用于间接计算 mRNA 的稳定性。通过这种动力学方法,我们还能够确定转录率和 mRNA 稳定性变化对偏离稳态的基因的 mRNA 变化的相对影响。

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