Nucleotide sequence analysis of in vivo recombinants between bacteriophage λ DNA and pBR322

King, Steven R.; Krolewski, M. A.; Marvo, Sandra L.; Lipson, P. J.; Pogue‐Geile, Kay L.; Chung, Jae Hoon; Jaskunas, S R

doi:10.1007/bf00337963

Cited by 13 publications

(11 citation statements)

References 39 publications

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“…Therefore, they suggested that the recombinations result from an exchange of DNA gyrase subunits bound to different sites. The nucleotide sequences at the A-plasmid junctions of one of these recombinants were determined and the results were consistent with this model (5).We have isolated several A-pBR322 recombinants that were generated in recA+ and recA-derivatives of E. coli (6,9). The sequences at the A-pBR322 junctions of four of these recombinants were determined (6).…”

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confidence: 61%

“…Another possible explanation for some illegitimate recombination events is that they are generated by DNA gyrase (4)(5)(6). Ikeda et aL (4,5) discovered that illegitimate recombinations between A DNA and plasmid pBR322 occur in extracts of E. coli and that the frequency of recombination is increased by the addition of oxolinic acid (4,5), which is an inhibitor of DNA gyrase (7,8).…”

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confidence: 99%

“…We have isolated several A-pBR322 recombinants that were generated in recA+ and recA-derivatives of E. coli (6,9). The sequences at the A-pBR322 junctions of four of these recombinants were determined (6).…”

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confidence: 99%

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Role of short regions of homology in intermolecular illegitimate recombination events.

Marvo¹,

King²,

Jaskunas³

1983

Proc. Natl. Acad. Sci. U.S.A.

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The structures of three recombinants between bacteriophage A DNA and plasmid pBR322 that were generated in a recA derivative of Escherichia coli are described. Each resulted from two illegitimate recombination events that resulted in the substitution of part of the A genome by part of the plasmid genome. The nucleotide sequences at the six A-plasmid junctions were determined and compared with the sequences of the A and plasmid genomes before recombination. Each recombination occurred at a short region of homology in the two genomes, and other short regions of homology were found near some of the junctions. The structures of these junctions are similar to those resulting from illegitimate recombination in animal cells. A model to explain how these multiple illegitimate recombination events could result from a cascade of DNA gyrase-catalyzed recombinations is discussed.The mechanisms responsible for generating gross DNA rearrangements such as deletions, substitutions, inversions, amplifications, and translocations are not understood. Recently, two specific models have been proposed that could account for these illegitimate recombination events.Miller and co-workers (1, 2) have found that most deletions in Escherichia coli occur between directly repeated sequences of 5 or more base pairs (bp). Therefore, they suggested that these deletions resulted from a slippage error in replication (2). Efstratiadis et al. (3) have suggested a similar model for the generation of deletions in eukaryotic cells.Another possible explanation for some illegitimate recombination events is that they are generated by DNA gyrase (4-6). Ikeda et aL (4, 5) discovered that illegitimate recombinations between A DNA and plasmid pBR322 occur in extracts of E. coli and that the frequency of recombination is increased by the addition of oxolinic acid (4, 5), which is an inhibitor of DNA gyrase (7,8). Therefore, they suggested that the recombinations result from an exchange of DNA gyrase subunits bound to different sites. The nucleotide sequences at the A-plasmid junctions of one of these recombinants were determined and the results were consistent with this model (5).We have isolated several A-pBR322 recombinants that were generated in recA+ and recA-derivatives of E. coli (6,9). The sequences at the A-pBR322 junctions of four of these recombinants were determined (6). Each resulted from reciprocal recombination between 10-13 bp of homology in A DNA and pBR322, including one (ATA1R) that was isolated in a recAstrain. One of the recombinants, AKA7, had an unusual structure in that it resulted from two of these rare events. DNA gyrase appears to be capable of catalyzing multiple recombinations (4). Therefore, we have suggested that AKA7 may have been generated by a cascade of DNA gyrase-catalyzed recombinations (6).The structures of three additional A-pBR322 recombinants isolated in a recA-strain of E. coli are described here. Each resulted from two recombinations at sites of 1-5 bp of homology in the A and plasmid genomes. Their structur...

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confidence: 61%

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confidence: 99%

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Role of short regions of homology in intermolecular illegitimate recombination events.

Marvo¹,

King²,

Jaskunas³

1983

Proc. Natl. Acad. Sci. U.S.A.

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“…The integration products of pE194 recombination show similarity to the products of other illegitimate recombinations, such as the recA-independent cointegrate fusions between pBR322 and the F-factor oriVI region (33) and also lambda-pBR322 recombination (24,27). These recombination events occur at low frequency (10-8 to 10-9) in E. coli recA mutants, and the recombination products appear to rise from reciprocal recombination between sequence identities of 10 to 13 bp (24,33).…”

Section: Discussionmentioning

confidence: 88%

“…These recombination events occur at low frequency (10-8 to 10-9) in E. coli recA mutants, and the recombination products appear to rise from reciprocal recombination between sequence identities of 10 to 13 bp (24,33). Most models for producing illegitimate recombination events of this type involve errors in DNA metabolism at first proposed by Franklin (10).…”

Section: Discussionmentioning

confidence: 99%

Identification of plasmid and Bacillus subtilis chromosomal recombination sites used for pE194 integration

Dempsey¹,

Dubnau²

1989

J Bacteriol

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The plasmid pE194 (3.7 kilobases) is capable of integrating into the genome of the bacterial host Bacillus subtilis in the absence of the major homology-dependent RecE recombination system. Multiple recombination sites have been identified on both the B. subtilis chromosome and pE194 (J. Hofemeister, M. Israeli-Reches, and D. Dubnau, Mol. Gen. Genet. 189:58-68, 1983). The B. subtilis chromosomal recombination sites were recovered by genetic cloning, and these sites were studied by nucleotide sequence analysis. Recombination had occurred between regions of short nucleotide homology (6 to 14 base pairs) as indicated by comparison of the plasmid and the host chromosome recombination sites with the crossover sites of the integration products. Recombination between the homologous sequences of the plasmid and the B. subtilis genome produced an integrated pE194 molecule which was bounded by direct repeats of the short homology. These results suggest a recombination model involving a conservative, reciprocal strand exchange between the two recombination sites. A preferred plasmid recombination site was found to occur within a 70-base-pair region which contains a GC-rich dyad symmetry element. Five of seven pE194-integrated strains analyzed had been produced by recombination at different locations within this 70-base-pair interval, located between positions 860 and 930 in pE194. On the basis of these data, mechanisms are discussed to explain the recombinational integration of pE194.Several intramolecular and intermolecular genetic recombination events which do not require extensive nucleotide sequence homology have been described for Bacillus subtilis (18,23,25,34). These rare events are independent of the major homologous DNA recombination system, the RecE pathway, of B. subtilis and produce stable rearrangements in the cellular genetic material. Often these genetic rearrangements are accompanied by changes in the phenotype of the host cell and provide a means of evolving new traits, such as the stable acquisition of antibiotic resistance determinants. The integration of plasmid pE194 into the B. subtilis genome (19) represents an example of recE-independent intermolecular recombination. Integration of pE194 places the ermC antibiotic resistance determinant into the B. subtilis chromosome. This gene is subsequently inherited as a chromosomal marker.pE194 is a 3.7-kilobase (kb) multicopy plasmid, originally isolated from Staphylococcus alureus (22), that was stably introduced into B. slubtilis by genetic transformation (13). The entire nucleotide sequence is known for pE194 (20). The ermC gene encodes an inducible 29-kilodalton product which is responsible for mediating resistance to the macrolidelincosamide-streptogramin B (MLS) group of antibiotics (45). pE194 is known to be naturally temperature sensitive for replication (39), being unable to replicate autonomously at growth temperatures exceeding 43°C. The plasmid-encoded trans-acting RepF protein is required for initiation of pE194 replication (16,44), which is tho...

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Transitory recombination between plasmid pHV33 and phage M13.

Dagert¹,

Ehrlich²

1983

The EMBO Journal

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Nucleotide sequence analysis of in vivo recombinants between bacteriophage λ DNA and pBR322

Cited by 13 publications

References 39 publications

Role of short regions of homology in intermolecular illegitimate recombination events.

Role of short regions of homology in intermolecular illegitimate recombination events.

Identification of plasmid and Bacillus subtilis chromosomal recombination sites used for pE194 integration

Transitory recombination between plasmid pHV33 and phage M13.

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