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Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP: Analysis of gene expression during potato tuber development
Author(s) -
Bachem Christian W.B.,
Hoeven Rutger S.,
Bruijn Steef M.,
Vreugdenhil Dick,
Zabeau Marc,
Visser Richard G.F.
Publication year - 1996
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1046/j.1365-313x.1996.9050745.x
Subject(s) - amplified fragment length polymorphism , complementary dna , gene , biology , gene expression , genetics , rna , coding region , microbiology and biotechnology , demography , genetic diversity , population , sociology
Summary Using a highly synchronous in vitro tuberization system, in combination with an amplified restriction fragment polymorphism (AFLP † )‐derived technique for RNA fingerprinting (cDNA‐AFLP ‡ ), transcriptional changes at and around the time point of potato tuberization have been analyzed. The targeted expression analysis of a specific transcript coding for the major potato storage protein, patatin and a second transcript, coding for ADP‐glucose pyrophosphorylase, a key gene in the starch biosynthetic pathway is described. This paper confirms that kinetics of expression revealed by cDNA‐AFLP analysis are comparable to those found in Northern analysis. Furthermore, this paper reports the isolation and analysis of two tuberspecific transcript‐derived fragments (TDFs) coding for the lipoxygenase enzyme, which are differentially induced around the time point of tuber formation. Analysis of the two lox TDFs demonstrates that it is possible to dissect the expression modalities of individual transcripts, not independently detectable by Northern analysis. Finally, it is shown that using cDNA‐AFLP, rapid and simple verification of band identity may be achieved. The results indicate that cDNA‐AFLP is a broadly applicable technology for identifying developmentally regulated genes.

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