Intramolecular transposition by a synthetic IS50 (Tn5) derivative.
We report the formation of deletions and inversions by intramolecular transposition of Tn5-derived mobile elements. The synthetic transposons used contained the IS50 O and I end segments and the transposase gene, a contraselectable gene encoding sucrose sensitivity (sacB), antibiotic resistance genes, and a plasmid replication origin. Both deletions and inversions were associated with loss of a 300-bp segment that is designated the vector because it is outside of the transposon. Deletions were severalfold more frequent than inversions, perhaps reflecting constraints on DNA twisting or abortive transposition. Restriction and DNA sequence analyses showed that both types of rearrangements extended from one transposon end to many different sites in target DNA. In the case of inversions, transposition generated 9-bp direct repeats of target sequences.
Tomcsanyi
T
T
Department of Molecular Microbiology, Washington University Medical School, St. Louis, Missouri 63110-1093.
Berg
C M
CM
Phadnis
S H
SH
Berg
D E
DE
D001483:Base Sequence; D002872:Chromosome Deletion; D007446:Chromosome Inversion; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005798:Genes, Bacterial; D005813:Genes, Synthetic; D008969:Molecular Sequence Data; D009713:Nucleotidyltransferases / Q000235:genetics*; D015345:Oligonucleotide Probes; D010957:Plasmids; D015183:Restriction Mapping; D013395:Sucrose / Q000378:metabolism; D019895:Transposases
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D015345:Oligonucleotide Probes; D013395:Sucrose; D009713:Nucleotidyltransferases; D019895:Transposases