Characterization and mapping of Ds-GUS-T-DNA lines for targeted insertional mutagenesis.
The transposition patterns of the Ds-GUS transposon T-DNA in 23 independent single-copy lines have been characterized and the map positions of 10 of them on three of the five Arabidopsis chromosomes are reported. Using overexpressed Activator (Ac) elements as a transposase source, it was found that the primary determinant of transposition frequency is the insertion site of the Ac-T-DNA. Neither the structure of the transposon T-DNA nor, in most cases, its insertion site have a significant effect on transposition frequency. Both the frequency and timing of transposition are influenced by the parent through which the transposon and transposase T-DNAs are transmitted. Overall, nearly 75% of plants in which excision has occurred bear a reinserted element and very short-range transpositions predominate, underlining the advantage of using mapped transposons for insertional mutagenesis.
Carnegie Institution of Washington, Department of Embryology, Baltimore, MD 21210, USA.
D
D
Smith
Y
Y
Yanai
Y G
YG
Liu
S
S
Ishiguro
K
K
Okada
D
D
Shibata
R F
RF
Whittier
N V
NV
Fedoroff
D017360:Arabidopsis / Q000235:genetics*; D002874:Chromosome Mapping; D003433:Crosses, Genetic; D004254:DNA Nucleotidyltransferases / Q000235:genetics; D004251:DNA Transposable Elements / Q000235:genetics*; D018628:Gene Dosage; D017930:Genes, Reporter; D005819:Genetic Markers; D005854:Germ Cells; D005966:Glucuronidase / Q000235:genetics; D016254:Mutagenesis, Insertional / Q000379:methods*; D018517:Plant Roots / Q000254:growth & development; D011995:Recombination, Genetic; D019895:Transposases
D004251:DNA Transposable Elements; D005819:Genetic Markers; D004254:DNA Nucleotidyltransferases; D019895:Transposases; D005966:Glucuronidase