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An ordered Arabidopsis thaliana mitochondrial cDNA library on high‐density filters allows rapid systematic analysis of plant gene expression: a pilot study
Author(s) -
Giegé Philippe,
Konthur Zoltán,
Walter Gerald,
Brennicke Axel
Publication year - 1998
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.1998.00242.x
Subject(s) - biology , complementary dna , arabidopsis thaliana , gene , cdna library , computational biology , open reading frame , rapid amplification of cdna ends , genetics , arabidopsis , gene expression , genome , molecular cloning , peptide sequence , mutant
Summary The availability of the complete sequence of a genome allows a systematic analysis of its expression. Gene‐specific variations of transcription levels and phenomena such as transcript processing and RNA editing require large numbers of clones to be examined. For the completely sequenced mitochondrial genome of Arabidopsis thaliana we adapted robot technology to identify and characterize expressed genes. A cDNA library of about 50 000 clones was constructed, robot‐ordered into 384‐well microtitre plates and spotted onto high‐density filter membranes. These filters permit the isolation of large numbers of specific cDNA clones in a single hybridization step. The cox1 , cox2 and cox3 genes were used to evaluate the feasibility and efficiency of this approach. A cluster of RNA editing sites observed outside the cox3 coding region identifies a novel reading frame orf95 in higher plants with significant similarity to a subunit of respiratory chain complex II.