Characterization of the Ac/Ds behaviour in transgenic tomato plants using plasmid rescue.
We describe the use of plasmid rescue to facilitate studies on the behaviour of Ds and Ac elements in transgenic tomato plants. The rescue of Ds elements relies on the presence of a plasmid origin of replication and a marker gene selective in Escherichia coli within the element. The position within the genome of modified Ds elements, rescued both before and after transposition, is assigned to the RFLP map of tomato. Alternatively to the rescue of Ds elements equipped with plasmid sequences, Ac elements are rescued by virtue of plasmid sequences flanking the element. In this way, the consequences of the presence of an (active) Ac element on the DNA structure at the original site can be studied in detail. Analysis of a library of Ac elements, rescued from the genome of a primary transformant, shows that Ac elements are, infrequently, involved in the formation of deletions. In one case the deletion refers to a 174 bp genomic DNA sequence immediately flanking Ac. In another case, a 1878 bp internal Ac sequence is deleted.
Rommens
C M
CM
Free University, Department of Genetics, Amsterdam, Netherlands.
Rudenko
G N
GN
Dijkwel
P P
PP
van Haaren
M J
MJ
Ouwerkerk
P B
PB
Blok
K M
KM
Nijkamp
H J
HJ
Hille
J
J
D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004261:DNA Replication; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D010944:Plants / Q000235:genetics*; D030821:Plants, Genetically Modified; D010957:Plasmids; D012150:Polymorphism, Restriction Fragment Length; D015183:Restriction Mapping
D004251:DNA Transposable Elements; D004247:DNA