Summary
Illegitimate recombination dependent on T4 DNA topoisomerase in a cell-free system has recently been described. In that work, recombinants between two phage λ DNA molecules were produced by the topoisomerase alone, without an Escherichia coli extract. In this paper, it is shown that recombination between phage λ and circular plasmid DNA molecules can also be detected in the presence or absence of an E. coli extract but at frequencies two or three orders of magnitude lower than that observed in the phage-phage cross. The frequency is probably lower because multiple recombination is required in the case of the phage-plasmid cross.
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References
Botchan M, Stringer JR, Mitchison T, Sambrook J (1980) Integration and excision of SV40 DNA from the chromosome of a transformed cell. Cell 20:143–152
Ikeda H (1986) Bacteriophage T4 DNA topoisomerase mediates illegitimate recombination in vitro. Proc Natl Acad Sci USA, in press
Ikeda H, Shiozaki M (1984) Nonhomologous recombination mediated by Escherichia coli DNA gyrase: possible involvement of DNA replication. Cold Spring Harbor Symp Quant Biol 49:401–409
Ikeda H, Moriya K, Matsumoto T (1981) In vitro study of illegitimate recombination: Involvement of DNA gyrase. Cold Spring Harbor Symp Quant Biol 45:399–408
Ikeda H, Aoki K, Naito A (1982) Illegitimate recombination mediated in vitro by DNA gyrase of Escherichia coli: structure of recombinant DNA molecules. Proc Natl Acad Sci USA 79:3724–3728
Ikeda H, Kawasaki I, Gellert M (1984) Mechanism of illegitimate recombination: Common sites for recombination and cleavage mediated by E. coli DNA gyrase. Mol Gen Genet 196:546–549
Kreuzer KN, Jongeneel CV (1983) Escherichia coli phage T4 topoisomerase. Methods Enzymol 100:144–160
Liu LF, Liu C-C, Alberts BM (1979) T4 DNA topoisomerase: a new ATP-dependent enzyme essential for initiation of T4 bacteriophage DNA replication. Nature 281:456–461
Liu LF, Liu C-C, Alberts BM (1980) Type II DNA topoisomerases: Enzymes that can unknot a topologically knotted DNA molecule via a reversible double-strand break. Cell 19:697–707
Marvo SL, King SR, Jaskunas SR (1983) Role of short regions of homology in intermolecular illegitimate recombination events. Proc Natl Acad Sci USA 80:2452–2456
Naito S, Uchida H (1980) Initiation of DNA replication in a ColE1-type plasmid: Isolation of mutations in the ori region. Proc Natl Acad Sci USA 77:6744–6748
Naito A, Naito S, Ikeda H (1984) Homology is not required for recombination mediated by DNA gyrase of Escherichia coli. Mol Gen Genet 193:238–243
Seasholz AF, Greenberg GR (1983) Identification of bacteriophage T4 gene 60 product and a role for this protein in DNA topoisomerase. J Biol Chem 258:1221–1226
Stetler GL, King GJ, Huang WM (1979) T4 DNA-delay proteins, required for specific DNA replication, form a complex that has ATP-dependent DNA topoisomerase activity. Proc Natl Acad Sci USA 76:3737–3741
Stringer JR (1982) DNA sequence homology and chromosomal deletion at a site of SV40 DNA integration. Nature 296:363–366
Wold MS, Mallory JB, Roberts JD, LeBowitz JH, McMacken R (1982) Initiation of bacteriophage λ DNA replication in vitro with purified λ replication proteins. Proc Natl Acad Sci USA 79:6167–6180
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Communicated by M. Takanami
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Ikeda, H. Illegitimate recombination mediated by T4 DNA topoisomerase in vitro. Molec Gen Genet 202, 518–520 (1986). https://doi.org/10.1007/BF00333287
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DOI: https://doi.org/10.1007/BF00333287