Quantitative cDNA-AFLP analysis for genome-wide expression studies

被引：158

作者：

Breyne, P ^{[1
]}

Dreesen, R ^{[1
]}

Cannoot, B ^{[1
]}

Rombaut, D ^{[1
]}

Vandepoele, K ^{[1
]}

Rombauts, S ^{[1
]}

Vanderhaeghen, R ^{[1
]}

Inzé, D ^{[1
]}

Zabeau, M ^{[1
]}

机构：

[1] State Univ Ghent VIB, Depy Plant Syst Biol, B-9000 Ghent, Belgium

来源：

MOLECULAR GENETICS AND GENOMICS | 2003年 / 269卷 / 02期

关键词：

cell cycle; functional genomics; gene expression; microarrays; transcriptome; HISTONE GENE-EXPRESSION; CELL-CYCLE; DISPLAY;

D O I：

10.1007/s00438-003-0830-6

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

An improved cDNA-AFLP method for genome-wide expression analysis has been developed. We demonstrate that this method is an efficient tool for quantitative transcript profiling and a valid alternative to microarrays. Unique transcript tags, generated from reverse-transcribed messenger RNA by restriction enzymes, were screened through a series of selective PCR amplifications. Based on in silico analysis, an enzyme combination was chosen that ensures that at least 60% of all the mRNAs were represented by an informative sequence tag. The sensitivity and specificity of the method allows one to detect poorly expressed genes and distinguish between homologous sequences. Accurate gene expression profiles were determined by quantitative analysis of band intensities, and subtle differences in transcriptional activity were revealed. A detailed screen for cell cycle-modulated genes in tobacco demonstrates the usefulness of the technology for genome-wide expression analysis.

引用

页码：173 / 179

页数：7

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