High rates of Ac/Ds germinal transposition in Arabidopsis suitable for gene isolation by insertional mutagenesis.
Overexpression of the Activator (Ac) transposase gene in Arabidopsis thaliana resulted in a minimal germinal transposition frequency of 27% in which independent Dissociation (Ds) transposition events were observed. Molecular analysis of 45 F1 generation Ac/Ds plants indicated that high rates of somatic excision had occurred, and independent germinal insertions were identified in F2 generation progeny plants. A tandem cauliflower mosaic virus (CaMV) promoter fused to two different Ac coding sequences significantly increased the rate of Ds transposition. The CaMV-Ac fusions activated single and multiple copies of two different Ds elements, DsDHFR and Ds35S-1, and reciprocal crosses resulted in similar transposition frequencies. The improved rate of independent germinal transposition observed makes Arabidopsis an ideal system for insertional mutagenesis.
Grevelding
C
C
Max-Planck-Institut für Züchtungsforschung, Cologne, Federal Republic of Germany.
Becker
D
D
Kunze
R
R
von Menges
A
A
Fantes
V
V
Schell
J
J
Masterson
R
R
D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D015870:Gene Expression; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009713:Nucleotidyltransferases / Q000378:metabolism*; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D010944:Plants / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D012231:Rhizobium / Q000235:genetics; D019895:Transposases; D003313:Zea mays / Q000235:genetics
D004251:DNA Transposable Elements; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D009713:Nucleotidyltransferases; D019895:Transposases