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酿酒酵母中的全基因组表达监测。

Genome-wide expression monitoring in Saccharomyces cerevisiae.

作者信息

Wodicka L, Dong H, Mittmann M, Ho M H, Lockhart D J

机构信息

Affymetrix, Santa Clara, CA 95051, USA.

出版信息

Nat Biotechnol. 1997 Dec;15(13):1359-67. doi: 10.1038/nbt1297-1359.

Abstract

The genomic sequence of the budding yeast Saccharomyces cerevisiae has been used to design and synthesize high-density oligonucleotide arrays for monitoring the expression levels of nearly all yeast genes. This direct and highly parallel approach involves the hybridization of total mRNA populations to a set of four arrays that contain a total of more than 260,000 specifically chosen oligonucleotides synthesized in situ using light-directed combinatorial chemistry. The measurements are quantitative, sensitive, specific, and reproducible. Expression levels ranging from less than 0.1 copies to several hundred copies per cell have been measured for cells grown in rich and minimal media. Nearly 90% of all yeast mRNAs are observed to be present under both conditions, with approximately 50% present at levels between 0.1 and 1 copy per cell. Many of the genes observed to be differentially expressed under these conditions are expected, but large differences are also observed for many previously uncharacterized genes.

摘要

芽殖酵母酿酒酵母的基因组序列已被用于设计和合成高密度寡核苷酸阵列,以监测几乎所有酵母基因的表达水平。这种直接且高度并行的方法涉及将总mRNA群体与一组四个阵列进行杂交,这些阵列总共包含超过260,000个使用光导向组合化学原位合成的特定选择的寡核苷酸。这些测量是定量的、灵敏的、特异的且可重复的。对于在丰富和基本培养基中生长的细胞,已测量出每个细胞中表达水平从少于0.1个拷贝到数百个拷贝不等。在这两种条件下,几乎所有酵母mRNA的90%都被观察到存在,其中约50%以每个细胞0.1至1个拷贝的水平存在。在这些条件下观察到差异表达的许多基因是预期的,但对于许多以前未表征的基因也观察到了很大差异。

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