Summary
Bovine papillomavirus (BPV) shuttle vectors replicate as a circular plasmid in mouse cell nuclei without impairing host cell viability. We used these vectors to analyze homologous recombination in mammalian cells. When several BPV-based plasmids carrying direct repeats were introduced into C127 cells, we detected many recombinant plasmid molecules that have lost the sequence between the repeats. Many recombinant type molecules as well as parental type molecules were detected in all the cell clones isolated for analysis. Sequencing after rescue of the plasmid inEscherichia coli showed that most of the recombinants were from accurate homologous recombination. When the repeats on the plasmid were in inverted orientation, no crossing-over type products were detected. We discuss possible mechanisms that explain these features.
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References
Albertini AM, Hofer M, Calos MP, Miller JH (1982) On the formation of spontaneous deletions: the importance of short sequence homologies in the generation of large deletions. Cell 29:319–328
Bachman AR, Burnett L, Berg P (1978) The SV40 nucleotide sequence. In: Tooze J (ed) DNA tumor viruses. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 791–841
Bollag RJ, Liskay RM (1988) Conservative intrachromosomal recombination between inverted repeats in mouse cells: association between reciprocal exchange and gene conversion. Genetics 119:161–169
Brouillette S, Chartrand P (1987) Intermolecular recombination asay for mammalian cells that produces recombinants carrying both homologous and nonhomologous junction. Mol Cell Biol 7:2248–2255
Chakrabarti S, Seidman MM (1986) Intermolecular recombination between transfected repeated sequences in mammalian cells is nonconservative. Mol Cell Biol 6:2520–2526
Church GM, Gilbert W (1984) Genomic sequencing. Proc Natl Acad Sci USA 81:1991–1995
Craigie R, Mizuuchi K (1986) Role of DNA topology in Mu transposition: mechanism of sensing the relative orientation of two DNA segments. Cell 45:793–800
DuBridge RB, Tang P, Hsia HC, Leong P-M, Miller JH, Calos M (1987) Analysis of mutation in human cells by using an Epstein-Barr virus shuttle system. Mol Cell Biol 7:379–387
Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specificity. Anal Biochem 132:6–13
Graham FL, van der Eb AJ (1973) A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52:456–467
Hanahan D (1983) Studies on transformation ofEscherichia coli with plasmids. J Mol Biol 166:557–580
Hirt B (1967) Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol 26:365–368
Klar AJS (1987) Differentiated parental DNA strands confer developmental asymmetry on daughter cells in fission yeast. Nature 326:466–470
Kobayashi I (1991) Mechanisms for gene conversion and homologous recombination: The double-strand break repair model and the successive half crossing-over model. Adv Biophys 28 (in press)
Krasnow MA, Cozzarelli NR (1983) Site-specific relaxation and recombination by Tn3 resolvase: recognition of the DNA path between oriented re sites. Cell 32:1313–1324
Lambert PL, Baker CC, Howley PM (1988) The genetics of bovine papillomavirus type 1. Annu Rev Genet 22:235–258
Lin F-L, Sperle K, Sternberg N (1984) Model for homologous recombination during transfer of DNA into mouse L cells: role for DNA ends in recombination process. Mol Cell Biol 4:1020–1034
Lin F-LM, Sperle KM, Sternberg NL (1987) Extrachromosomal recombination in mammalian cells as studied with single- and double-stranded DNA substrate. Mol Cell Biol 7:129–140
Liskay RM, Stachelek JL (1986) Information transfer between duplicated chromosomal sequences in mammalian cells involves contiguous regions of DNA. Proc Natl Acad Sci USA 83:1802–1806
Lusky M, Botchan M (1981) Inhibition of SV40 replication in simian cells by specific pBR322 DNA sequences. Nature 293:79–81
Lusky M, Botchan MR (1985) Genetic analysis of bovine papillomavirus type 1 trans-acting replication factors. J Virol 53:955–965
Parker BA, Stark GR (1979) Regulation of simian virus 40 transcription: sensitive analysis of RNA present early in infections by virus or virus DNA. J Virol 31:360–369
Rösl F, Waldeck W, Zentgraf H, Sauer G (1986) Properties of intracellular bovine papillomavirus chromatine. J Virol 58:500–507
Rubnitz J, Subramani S (1985) Rapid assay for extrachromosomal homologous recombination in monkey cells. Mol Cell Biol 5:529–537
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Sarver N, Byrne JC, Howley PM (1982) Transformation and replication in mouse cells of bovine papillomavirus/pML2 plasmid vectors that can be rescued in bacteria. Proc Natl Acad Sci USA 79:7147–7151
Smithies O, Gregg RG, Boggs SS, Koralewski MA, Kucherlapati RS (1985) Insertion of DNA sequences into the human chromosomal beta-globin locus by homologous recombination. Nature 317:230–234
Southern PJ, Berg P (1982) Transformation of mammalian cells to antibiotic resistance with bacterial gene under control of SV40 early region promoter. J Mol Appl Genet 1:327–341
Subramani S, Berg P (1983) Homologous and nonhomologous recombination in mammalian cells. Mol Cell Biol 3:1040–1052
Subramani S, Rubnitz J (1985) Recombination events after infection and stable integration of DNA into mouse cells. Mol Cell Biol 5:659–666
Subramani S, Seaton BL (1988) Homologous recombination in mitotically dividing mammalian cells. In: Kucherlapati R, Smith GR (eds) Genetic recombination. American Society for Microbiology, Washington DC, pp 549–573
Thomas KR, Capecchi MR (1986) Introduction of homologous DNA sequences into mammalian cells induces mutations in cognate gene. Nature 324:34–38
Thomas KR, Folger KR, Capecchi MR (1986) High frequency targeting of genes to specific site in the mammalian genome. Cell 44:419–428
Vieira J, Messing J (1987) Production of single-stranded plasmid DNA. Methods Enzymol 153:3–11
Wake CT, Vernaleone F, Wilson JH (1985) Topological requirements for homologous recombination among DNA molecules transfected into mammalian cells. Mol Cell Biol 5:2080–2089
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Communicated by M. Sekiguchi
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Kitamura, Y., Yoshikura, H. & Kobayashi, I. Homologous recombination in a mammalian plasmid. Mol Gen Genet 222, 185–191 (1990). https://doi.org/10.1007/BF00633816
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DOI: https://doi.org/10.1007/BF00633816