Formation of deletion in Escherichia coli between direct repeats located in the long inverted repeats of a cellular slime mold plasmid: participation of DNA gyrase.
We constructed a recombinant plasmid containing the 2.1 kb HindIII fragment of plasmid pDG1, isolated from the cellular slime mold (Dictyostelium sp. strain GA11), and using pAG60 as cloning vector. We found that deletions of the recombinant plasmid took place frequently in Escherichia coli wild-type cells. However, the deletion was not observed when the plasmid was introduced into a strain that was an isogenic temperature-sensitive mutant of the gyrA gene. These results suggest that E. coli DNA gyrase is involved in the mechanisms of the deletion formation. It was shown that the 1.0 kb deletant derived from the 2.1 kb HindIII insert was produced by elimination of a 1.1 kb region. Sequence analysis of the deletants showed that cutting and rejoining took place between two out of the six nearly perfect direct repeats [21 bp palindromic sequences; AAAAAA(T/C)GGC(G/C)GCC(A/G)TTTTTT], located near the distal ends of the inverted repeats, preserving one copy of the repeats. These sequences consist of local short inverted repeats, where cutting and rejoining occur at one of the two regions.
Saing
K M
KM
Institute of Biological Sciences, University of Tsukuba, Ibaraki, Japan.
Orii
H
H
Tanaka
Y
Y
Yanagisawa
K
K
Miura
A
A
Ikeda
H
H
D001483:Base Sequence; D003001:Cloning, Molecular; D004250:DNA Topoisomerases, Type II / Q000378:metabolism*; D004271:DNA, Fungal; D004023:Dictyostelium / Q000235:genetics*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005796:Genes; D005798:Genes, Bacterial; D009154:Mutation; D010957:Plasmids; D015183:Restriction Mapping; D013696:Temperature
D004271:DNA, Fungal; D004250:DNA Topoisomerases, Type II