Abstract
In vivo recombination was found to occur between plasmid pHS1, a temperature-sensitive replication mutant of pSC101 carrying tetracycline resistance, and plasmid ColE1 after selection for tetracycline resistance at the restrictive temperature, 42 degrees C. Extensive analysis of the physical structures of three of these recombinant plasmids, using restriction endonucleases and the electron microscope heteroduplex method, revealed that the plasmid pHS1 was integrated into different sites on ColE1. The recombinant plasmids contained a duplication of a unique 1-kilobase (kb) sequence of pHS1 in a direct orientation at the junctions between the two parental plasmid sequences. This was confirmed by comparing the nucleotide sequence of the recombinants and their parental plasmids. Nucleotide sequence analysis further revealed that nine nucleotides at the site of recombination of ColE1 were duplicated at the junction of each of the 1-kb sequences. The formation of recombinants was independent of RecA function. Based on our previous finding that a plasmid containing a deoxyribonucleic acid insertion (IS) element can recombine with a second plasmid to generate a duplication of the IS element, we conclude that the 1-kb sequence is an insertion sequence, which we named IS102. For convenience, we have also denoted the IS102 sequence as eta theta to assign the orientation of the sequence. Eighteen nucleotides at one end (eta end) were found to be repeated in an inverted orientation at the other end (theta end) of IS102. The nucleotide sequence of the eta end of the sequence was found to be identical to the sequence at the ends of the transposon Tn903, which is responsible for transposition of the kanamycin resistance gene.
MeSH Terms
Base Sequence
DNA Replication
DNA Restriction Enzymes
DNA Transposable Elements
DNA, Bacterial/genetics
Escherichia coli/genetics
Nucleic Acid Heteroduplexes
Plasmids
Recombination, Genetic
Tetracycline/pharmacology
Chemicals
DNA Transposable Elements
DNA, Bacterial
Nucleic Acid Heteroduplexes
DNA Restriction Enzymes
Tetracycline
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Ohtsubo H
Zenilman M
Ohtsubo E
References (29)
29 references, click to expand
-
Origin of replication of colicin E1 plasmid DNA.
Proc Natl Acad Sci U S A. 1977 May;74(5):1865-9
PMID: 325558
-
Nucleotide-sequence analysis of Tn3 (ap): implications for insertion and deletion.
Cold Spring Harb Symp Quant Biol. 1979;43 Pt 2:1269-77
PMID: 385227
-
Isolation and characterization of transducing coliphage fd carrying a kanamycin resistance gene.
Gene. 1978 Feb;3(1):39-51
PMID: 344143
-
Plasmids containing insertion elements are potential transposons.
Proc Natl Acad Sci U S A. 1980 Feb;77(2):750-4
PMID: 6244582
-
Site specific recombination involved in the generation of small plasmids.
Mol Gen Genet. 1978 Feb 16;159(2):131-41
PMID: 345087
-
The transposon Tn9 generates a 9 bp repeated sequence during integration.
Cell. 1978 Dec;15(4):1209-19
PMID: 365355
-
Nucleotide sequence at the insertion sites of a kanamycin transposon.
Nature. 1978 Dec 21-28;276(5690):845-7
PMID: 723960
-
Replication of colicin E1 plasmid DNA in cell extracts. Origin and direction of replication.
Proc Natl Acad Sci U S A. 1974 Jun;71(6):2260-4
PMID: 4601821
-
Indirect selection of bacterial plasmids lacking identifiable phenotypic properties.
J Bacteriol. 1975 Oct;124(1):225-31
PMID: 1100600
-
Charon phages: safer derivatives of bacteriophage lambda for DNA cloning.
Science. 1977 Apr 8;196(4286):161-9
PMID: 847462
-
The use of thin acrylamide gels for DNA sequencing.
FEBS Lett. 1978 Mar 1;87(1):107-10
PMID: 631324
-
The ends of Tn10 are not IS3.
J Bacteriol. 1979 Sep;139(3):1097-101
PMID: 383689
-
Nucleotide sequence of bacteriophage phi X174 DNA.
Nature. 1977 Feb 24;265(5596):687-95
PMID: 870828
-
Electron microscope heteroduplex studies of sequence relations among plasmids of Escherichia coli. I. Structure of F-prime factors.
J Mol Biol. 1972 Nov 14;71(2):471-97
PMID: 4564488
-
Mutations of Bacteria from Virus Sensitivity to Virus Resistance.
Genetics. 1943 Nov;28(6):491-511
PMID: 17247100
-
Replication control in a composite plasmid constructed by in vitro linkage of two distinct replicons.
Nature. 1976 Jan 29;259(5541):285-90
PMID: 765836
-
Complete nucleotide sequence of the Escherichia coli plasmid pBR322.
Cold Spring Harb Symp Quant Biol. 1979;43 Pt 1:77-90
PMID: 383387
-
Mutations of temperature sensitivity in R plasmid pSC101.
J Bacteriol. 1977 Aug;131(2):405-12
PMID: 328479
-
Revised interpretation of the origin of the pSC101 plasmid.
J Bacteriol. 1977 Nov;132(2):734-7
PMID: 334752
-
The E. coli gene encoding heat stable toxin is a bacterial transposon flanked by inverted repeats of IS1.
Nature. 1979 Feb 8;277(5696):453-6
PMID: 368646
-
A second specific endonuclease from Haemophilus aegyptius.
J Mol Biol. 1975 Jan 5;91(1):121-3
PMID: 171410
-
Relaxation complexes of plasmids ColE1 and ColE2: unique site of the nick in the open circular DNA of the relaxed complexes.
Proc Natl Acad Sci U S A. 1974 Oct;71(10):3854-7
PMID: 4610563
-
Nucleotide sequence of an insertion element, IS1.
Proc Natl Acad Sci U S A. 1978 Feb;75(2):615-9
PMID: 273224
-
Electron microscope heteroduplex studies of sequence relations among plasmids of Escherichia coli. II. Structure of drug resistance (R) factors and F factors.
J Mol Biol. 1973 Apr 5;75(2):235-55
PMID: 4580676
-
The isolation and characterization of an in vivo recombinant between the filamentous bacteriophage f1 and the plasmid pSC101.
Virology. 1978 Sep;89(2):438-49
PMID: 362695
-
A new method for sequencing DNA.
Proc Natl Acad Sci U S A. 1977 Feb;74(2):560-4
PMID: 265521
-
Transposition of the Escherichia coli insertion element gamma generates a five-base-pair repeat.
Proc Natl Acad Sci U S A. 1979 Oct;76(10):4882-6
PMID: 388421
-
Replication of colicin E1 plasmid DNA in minicells from a unique replication initiation site.
Proc Natl Acad Sci U S A. 1974 Jun;71(6):2256-9
PMID: 4601583
-
Isolation, by tetracycline selection, of small plasmids derived from R-factor R12 in Escherichia coli K-12.
J Bacteriol. 1976 Jul;127(1):644-55
PMID: 776944