A novel transposon tagging element for obtaining gain-of-function mutants based on a self-stabilizing Ac derivative.
A novel tagging system AcREH, designed for obtaining gain-of-function mutations, was prepared on the basis of a self-stabilizing Ac transposon derivative. The transposable element, DsAT, was constructed in a way that it can activate transcription of neighboring genes by two 35S promoters and/or by four tandem repeats of the enhancer fragment of this promoter. DsAT revealed somatic excision in the first generation of the tobacco transformants. The element exhibited germinal excision to the next generation, as demonstrated by PCR and Southern hybridization analysis. In spite of the structure of the element, which may inhibit the expression of the transposase gene, the frequency of germinal excision was comparable to or higher than those so far reported, suggesting the applicability of the element for gene tagging.
Suzuki
Y
Y
Department of Applied Biological Chemistry, University of Tokyo, Japan. ayoshi@pgr1.ch.a.u-tokyo.ac.jp
Uemura
S
S
Saito
Y
Y
Murofushi
N
N
Schmitz
G
G
Theres
K
K
Yamaguchi
I
I
D001483:Base Sequence; D015139:Blotting, Southern; D017796:Caulimovirus / Q000235:genetics; D004251:DNA Transposable Elements / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D004274:DNA, Recombinant; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D016254:Mutagenesis, Insertional; D009154:Mutation; D010947:Plants, Toxic; D011401:Promoter Regions, Genetic / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D014026:Nicotiana / Q000235:genetics; D015533:Transcriptional Activation; D014170:Transformation, Genetic
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D018744:DNA, Plant; D004274:DNA, Recombinant; C036483:T-DNA