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. 1987 May;7(5):2007–2011. doi: 10.1128/mcb.7.5.2007

Recombination and ligation of transfected DNA in CHO mutant EM9, which has high levels of sister chromatid exchange.

C A Hoy, J C Fuscoe, L H Thompson
PMCID: PMC365311  PMID: 3600655

Abstract

Transformation frequencies were measured in CHO mutant EM9 after transfection with intact or modified plasmid pSV2-gpt. The mutant and wild-type strain behaved similarly under all conditions except when homologous recombination was required to produce an intact plasmid. Therefore, the defect of the mutant which renders it slow in DNA strand break rejoining and high in sister chromatid exchange induction reduces its ability to recombine foreign DNA molecules.

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Selected References

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  1. Ashman C. R., Davidson R. L. High spontaneous mutation frequency in shuttle vector sequences recovered from mammalian cellular DNA. Mol Cell Biol. 1984 Nov;4(11):2266–2272. doi: 10.1128/mcb.4.11.2266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brenner D. A., Smigocki A. C., Camerini-Otero R. D. Effect of insertions, deletions, and double-strand breaks on homologous recombination in mouse L cells. Mol Cell Biol. 1985 Apr;5(4):684–691. doi: 10.1128/mcb.5.4.684. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Calos M. P., Lebkowski J. S., Botchan M. R. High mutation frequency in DNA transfected into mammalian cells. Proc Natl Acad Sci U S A. 1983 May;80(10):3015–3019. doi: 10.1073/pnas.80.10.3015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chan J. Y., Thompson L. H., Becker F. F. DNA-ligase activities appear normal in the CHO mutant EM9. Mutat Res. 1984 May-Jun;131(5-6):209–214. doi: 10.1016/0167-8817(84)90027-0. [DOI] [PubMed] [Google Scholar]
  5. Cox R., Masson W. K., Debenham P. G., Webb M. B. The use of recombinant DNA plasmids for the determination of DNA-repair and recombination in cultured mammalian cells. Br J Cancer Suppl. 1984;6:67–72. [PMC free article] [PubMed] [Google Scholar]
  6. Fuscoe J. C., Fenwick R. G., Jr, Ledbetter D. H., Caskey C. T. Deletion and amplification of the HGPRT locus in Chinese hamster cells. Mol Cell Biol. 1983 Jun;3(6):1086–1096. doi: 10.1128/mcb.3.6.1086. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Giaccia A., Weinstein R., Hu J., Stamato T. D. Cell cycle-dependent repair of double-strand DNA breaks in a gamma-ray-sensitive Chinese hamster cell. Somat Cell Mol Genet. 1985 Sep;11(5):485–491. doi: 10.1007/BF01534842. [DOI] [PubMed] [Google Scholar]
  8. Hoy C. A., Salazar E. P., Thompson L. H. Rapid detection of DNA-damaging agents using repair-deficient CHO cells. Mutat Res. 1984 Oct;130(5):321–332. doi: 10.1016/0165-1161(84)90018-9. [DOI] [PubMed] [Google Scholar]
  9. Ikejima M., Bohannon D., Gill D. M., Thompson L. H. Poly(ADP-ribose) metabolism appears normal in EM9, a mutagen-sensitive mutant of CHO cells. Mutat Res. 1984 Sep;128(2):213–220. doi: 10.1016/0027-5107(84)90109-x. [DOI] [PubMed] [Google Scholar]
  10. Kemp L. M., Sedgwick S. G., Jeggo P. A. X-ray sensitive mutants of Chinese hamster ovary cells defective in double-strand break rejoining. Mutat Res. 1984 Nov-Dec;132(5-6):189–196. doi: 10.1016/0167-8817(84)90037-3. [DOI] [PubMed] [Google Scholar]
  11. Kucherlapati R. S., Eves E. M., Song K. Y., Morse B. S., Smithies O. Homologous recombination between plasmids in mammalian cells can be enhanced by treatment of input DNA. Proc Natl Acad Sci U S A. 1984 May;81(10):3153–3157. doi: 10.1073/pnas.81.10.3153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Latt S. A. Sister chromatid exchange formation. Annu Rev Genet. 1981;15:11–55. doi: 10.1146/annurev.ge.15.120181.000303. [DOI] [PubMed] [Google Scholar]
  13. Lin F. L., Sperle K., Sternberg N. Model for homologous recombination during transfer of DNA into mouse L cells: role for DNA ends in the recombination process. Mol Cell Biol. 1984 Jun;4(6):1020–1034. doi: 10.1128/mcb.4.6.1020. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Miller C. K., Temin H. M. High-efficiency ligation and recombination of DNA fragments by vertebrate cells. Science. 1983 May 6;220(4597):606–609. doi: 10.1126/science.6301012. [DOI] [PubMed] [Google Scholar]
  15. Mulligan R. C., Berg P. Factors governing the expression of a bacterial gene in mammalian cells. Mol Cell Biol. 1981 May;1(5):449–459. doi: 10.1128/mcb.1.5.449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mulligan R. C., Berg P. Selection for animal cells that express the Escherichia coli gene coding for xanthine-guanine phosphoribosyltransferase. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2072–2076. doi: 10.1073/pnas.78.4.2072. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pinkel D., Thompson L. H., Gray J. W., Vanderlaan M. Measurement of sister chromatid exchanges at very low bromodeoxyuridine substitution levels using a monoclonal antibody in Chinese hamster ovary cells. Cancer Res. 1985 Nov;45(11 Pt 2):5795–5798. [PubMed] [Google Scholar]
  18. Pomerantz B. J., Naujokas M., Hassell J. A. Homologous recombination between transfected DNAs. Mol Cell Biol. 1983 Sep;3(9):1680–1685. doi: 10.1128/mcb.3.9.1680. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Radding C. M. Homologous pairing and strand exchange in genetic recombination. Annu Rev Genet. 1982;16:405–437. doi: 10.1146/annurev.ge.16.120182.002201. [DOI] [PubMed] [Google Scholar]
  20. Razzaque A., Mizusawa H., Seidman M. M. Rearrangement and mutagenesis of a shuttle vector plasmid after passage in mammalian cells. Proc Natl Acad Sci U S A. 1983 May;80(10):3010–3014. doi: 10.1073/pnas.80.10.3010. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  22. Roberstson M. Gene rearrangement and the generation of diversity. Nature. 1982 May 20;297(5863):184–186. doi: 10.1038/297184a0. [DOI] [PubMed] [Google Scholar]
  23. Shimizu A., Honjo T. Immunoglobulin class switching. Cell. 1984 Apr;36(4):801–803. doi: 10.1016/0092-8674(84)90029-1. [DOI] [PubMed] [Google Scholar]
  24. Small J., Scangos G. Recombination during gene transfer into mouse cells can restore the function of deleted genes. Science. 1983 Jan 14;219(4581):174–176. doi: 10.1126/science.6294829. [DOI] [PubMed] [Google Scholar]
  25. Song K. Y., Chekuri L., Rauth S., Ehrlich S., Kucherlapati R. Effect of double-strand breaks on homologous recombination in mammalian cells and extracts. Mol Cell Biol. 1985 Dec;5(12):3331–3336. doi: 10.1128/mcb.5.12.3331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  27. Szostak J. W., Orr-Weaver T. L., Rothstein R. J., Stahl F. W. The double-strand-break repair model for recombination. Cell. 1983 May;33(1):25–35. doi: 10.1016/0092-8674(83)90331-8. [DOI] [PubMed] [Google Scholar]
  28. Thompson L. H., Brookman K. W., Dillehay L. E., Carrano A. V., Mazrimas J. A., Mooney C. L., Minkler J. L. A CHO-cell strain having hypersensitivity to mutagens, a defect in DNA strand-break repair, and an extraordinary baseline frequency of sister-chromatid exchange. Mutat Res. 1982 Aug;95(2-3):427–440. doi: 10.1016/0027-5107(82)90276-7. [DOI] [PubMed] [Google Scholar]
  29. Thompson L. H., Brookman K. W., Dillehay L. E., Mooney C. L., Carrano A. V. Hypersensitivity to mutation and sister-chromatid-exchange induction in CHO cell mutants defective in incising DNA containing UV lesions. Somatic Cell Genet. 1982 Nov;8(6):759–773. doi: 10.1007/BF01543017. [DOI] [PubMed] [Google Scholar]
  30. Thompson L. H., Brookman K. W., Minkler J. L., Fuscoe J. C., Henning K. A., Carrano A. V. DNA-mediated transfer of a human DNA repair gene that controls sister chromatid exchange. Mol Cell Biol. 1985 Apr;5(4):881–884. doi: 10.1128/mcb.5.4.881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Thompson L. H., Fong S., Brookman K. Validation of conditions for efficient detection of HPRT and APRT mutations in suspension-cultured Chinese hamster ovary cells. Mutat Res. 1980 Feb;74(1):21–36. doi: 10.1016/0165-1161(80)90188-0. [DOI] [PubMed] [Google Scholar]
  32. Thompson L. H., Rubin J. S., Cleaver J. E., Whitmore G. F., Brookman K. A screening method for isolating DNA repair-deficient mutants of CHO cells. Somatic Cell Genet. 1980 May;6(3):391–405. doi: 10.1007/BF01542791. [DOI] [PubMed] [Google Scholar]
  33. Upcroft P., Carter B., Kidson C. Analysis of recombination in mammalian cells using SV40 genome segments having homologous overlapping termini. Nucleic Acids Res. 1980 Jun 25;8(12):2725–2736. doi: 10.1093/nar/8.12.2725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Volkert F. C., Young C. S. The genetic analysis of recombination using adenovirus overlapping terminal DNA fragments. Virology. 1983 Feb;125(1):175–193. doi: 10.1016/0042-6822(83)90072-7. [DOI] [PubMed] [Google Scholar]
  35. Wake C. T., Wilson J. H. Simian virus 40 recombinants are produced at high frequency during infection with genetically mixed oligomeric DNA. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2876–2880. doi: 10.1073/pnas.76.6.2876. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Yunis J. J. The chromosomal basis of human neoplasia. Science. 1983 Jul 15;221(4607):227–236. doi: 10.1126/science.6336310. [DOI] [PubMed] [Google Scholar]
  37. de Saint Vincent B. R., Wahl G. M. Homologous recombination in mammalian cells mediates formation of a functional gene from two overlapping gene fragments. Proc Natl Acad Sci U S A. 1983 Apr;80(7):2002–2006. doi: 10.1073/pnas.80.7.2002. [DOI] [PMC free article] [PubMed] [Google Scholar]

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