Identification of transferred DNA insertions within Arabidopsis genes involved in signal transduction and ion transport

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 93; no. 15; pp. 8145 - 8150
• Main Authors: Krysan, P.J. (University of Wisconsin, Madison, WI.), Young, J.C, Tax, F, Sussman, M.R
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 23.07.1996; National Acad Sciences; National Academy of Sciences
• Subjects: ADN; AGROBACTERIUM TUMEFACIENS; Agrobacterium tumefaciens - genetics; Arabidopsis - genetics; Arabidopsis Proteins; ARABIDOPSIS THALIANA; Base Sequence; Biological Transport; Biology; Botany; Calcium-Calmodulin-Dependent Protein Kinases - genetics; Cell Line, Transformed; Deoxyribonucleic acid; DNA; DNA Primers; DNA Transposable Elements; DNA, Bacterial - genetics; DNA, Bacterial - metabolism; DNA, Plant - analysis; DNA, Plant - genetics; DNA, Single-Stranded - genetics; DNA, Single-Stranded - metabolism; ENSAYO; FISIOLOGIA VEGETAL; Flowers & plants; GENE; Gene Deletion; GENES; Genes, Plant; Genetic screening; GENETICA; GENETIQUE; Genomics; ION; Ion transport; IONES; Multigene Family; MUTACION; Mutagenesis, Insertional; MUTATION; Nucleotide sequences; PCR; PHYSIOLOGIE VEGETALE; Plants; Polymerase Chain Reaction; PROTEINA QUINASA; PROTEINE KINASE; Reverse genetics; Signal Transduction; TESTAGE
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.93.15.8145

The transferred DNA (T-DNA) of Agrobacterium tumefaciens serves as an insertional mutagen once integrated into a host plant's genome. As a means of facilitating reverse genetic analysis in Arabidopsis thaliana, we have developed a method that allows one to search for plants carrying T-DNA insertions within any sequenced Arabidopsis gene. Using PCR, we screened a collection of 9100 independent T-DNA-transformed Arabidopsis lines and found 17 T-DNA insertions within the 63 genes analyzed. The genes surveyed include members of various gene families involved in signal transduction and ion transport. As an example, data are shown for a T-DNA insertion that was found within CPK-9, a member of the gene family encoding calmodulin-domain protein kinases