Transactivation of Ds by Ac-transposase gene fusions in tobacco.
To study regulation of the (Ds) transposition process in heterologous plant species, the transposase gene of Ac was fused to several promoters that are active late during plant development. These promoters are the flower-specific chalcone synthase A promoter (CHS A), the anther-specific chalcone isomerase B promoter CHI B and the pollen-specific chalcone isomerase A2 promoter CHI A2. The modified transposase genes were introduced into a tobacco tester plant. This plant contains Ds stably inserted within the leader sequence of the hygromycin resistance (HPT II) gene. As confirmed with positive control elements, excision of Ds leads to the restoration of a functional HPT II gene and to a hygromycin resistant phenotype. No hygromycin resistance was observed in negative control experiments with Ac derivatives lacking 5' regulatory sequences. Although transactivation of Ds was observed after the introduction of transposase gene fusions in calli, excision in regenerated plants was observed only for the CHS A- or CHI B-transposase gene fusions. With these modified transposase genes, somatic excision frequencies were increased (68%) and decreased (22%), respectively, compared to the situation with the Ac element itself (38%). The shifts in transactivation frequencies were not associated with significant differences in the frequencies of germinally transmitted excision events (approximately 5%). The relative somatic stability of Ds insertions bearing the CHI B-transposase gene fusion suggests the usefulness of this activator element for transposon tagging experiments.
Rommens
C M
CM
Free University, Department of Genetics, Amsterdam, The Netherlands.
van Haaren
M J
MJ
Buchel
A S
AS
Mol
J N
JN
van Tunen
A J
AJ
Nijkamp
H J
HJ
Hille
J
J
D000217:Acyltransferases / Q000235:genetics; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004251:DNA Transposable Elements; D019753:Intramolecular Lyases; D007535:Isomerases / Q000235:genetics; D008969:Molecular Sequence Data; D009713:Nucleotidyltransferases / Q000235:genetics*; D009838:Oligodeoxyribonucleotides; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D021382:Protein Sorting Signals / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping; D014026:Nicotiana / Q000201:enzymology / Q000235:genetics*; D019895:Transposases
D004251:DNA Transposable Elements; D009838:Oligodeoxyribonucleotides; D021382:Protein Sorting Signals; D004247:DNA; D000217:Acyltransferases; C020141:flavanone synthetase; D009713:Nucleotidyltransferases; D019895:Transposases; D007535:Isomerases; D019753:Intramolecular Lyases; C019865:chalcone isomerase