Identification of transferred DNA insertions within Arabidopsis genes involved in signal transduction and ion transport.
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 F-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.
Krysan
P J
PJ
Cell and Molecular Biology Program and the Department of Horticulture, University of Wisconsin, Madison 53706, USA.
Young
J C
JC
Tax
F
F
Sussman
M R
MR
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D017360:Arabidopsis / Q000235:genetics*; D029681:Arabidopsis Proteins; D001483:Base Sequence; D001692:Biological Transport; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000235:genetics*; D002461:Cell Line, Transformed; D017931:DNA Primers; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism*; D018744:DNA, Plant / Q000032:analysis / Q000235:genetics; D004277:DNA, Single-Stranded / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D017343:Genes, Plant; D005810:Multigene Family; D016254:Mutagenesis, Insertional; D016133:Polymerase Chain Reaction; D015398:Signal Transduction
D029681:Arabidopsis Proteins; D017931:DNA Primers; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D018744:DNA, Plant; D004277:DNA, Single-Stranded; C036483:T-DNA; C101493:CPK9 protein, Arabidopsis; D017871:Calcium-Calmodulin-Dependent Protein Kinases