Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1984 Nov;4(11):2266–2272. doi: 10.1128/mcb.4.11.2266

High spontaneous mutation frequency in shuttle vector sequences recovered from mammalian cellular DNA.

C R Ashman, R L Davidson
PMCID: PMC369054  PMID: 6096690

Abstract

The recombinant shuttle vector pSV2gpt was introduced into V79 Chinese hamster cells, and stable transformants expressing the Escherichia coli gpt gene were selected. Two transformants carrying tandem duplications of the plasmid at a single site were identified and fused to simian COS-1 cells. Plasmid DNA recovered from the heterokaryons was used to transform a Gpt- derivative of E. coli HB101, and the relative frequency of plasmids carrying a mutation in the gpt gene was determined. The high frequency of Gpt- plasmids (ca. 1%) was similar to that observed when plasmid was recovered from COS-1 cells which had been transfected with pSV2gpt. Most of the mutant plasmids had rearrangements in the region containing the gpt gene.

Full text

PDF
2266

Images in this article

2268Image
on p.2268
2268Image
on p.2268
2268Image
on p.2268
2269Image
on p.2269
2270Image
on p.2270
2270Image
on p.2270

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blin N., Stafford D. W. A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Res. 1976 Sep;3(9):2303–2308. doi: 10.1093/nar/3.9.2303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Breitman M. L., Tsui L. C., Buchwald M., Siminovitch L. Introduction and recovery of a selectable bacterial gene from the genome of mammalian cells. Mol Cell Biol. 1982 Aug;2(8):966–976. doi: 10.1128/mcb.2.8.966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Clough D. W., Kunkel L. M., Davidson R. L. 5-Azacytidine-induced reactivation of a herpes simplex thymidine kinase gene. Science. 1982 Apr 2;216(4541):70–73. doi: 10.1126/science.6175023. [DOI] [PubMed] [Google Scholar]
  6. Conrad S. E., Liu C. P., Botchan M. R. Fragment spanning the SV40 replication origin is the only DNA sequence required in cis for viral excision. Science. 1982 Dec 17;218(4578):1223–1225. doi: 10.1126/science.6293055. [DOI] [PubMed] [Google Scholar]
  7. Davidson R. L., Adelstein S. J., Oxman M. N. Herpes simplex virus as a source of thymidine kinase for thymidine kinase-deficient mouse cells: suppression and reactivation of the viral enzyme. Proc Natl Acad Sci U S A. 1973 Jul;70(7):1912–1916. doi: 10.1073/pnas.70.7.1912. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Davies R. L., Fuhrer-Krusi S., Kucherlapati R. S. Modulation of transfected gene expression mediated by changes in chromatin structure. Cell. 1982 Dec;31(3 Pt 2):521–529. doi: 10.1016/0092-8674(82)90308-7. [DOI] [PubMed] [Google Scholar]
  9. FORD D. K., YERGANIAN G. Observations on the chromosomes of Chinese hamster cells in tissue culture. J Natl Cancer Inst. 1958 Aug;21(2):393–425. [PubMed] [Google Scholar]
  10. Gluzman Y. SV40-transformed simian cells support the replication of early SV40 mutants. Cell. 1981 Jan;23(1):175–182. doi: 10.1016/0092-8674(81)90282-8. [DOI] [PubMed] [Google Scholar]
  11. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  12. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  13. LITTLEFIELD J. W. SELECTION OF HYBRIDS FROM MATINGS OF FIBROBLASTS IN VITRO AND THEIR PRESUMED RECOMBINANTS. Science. 1964 Aug 14;145(3633):709–710. doi: 10.1126/science.145.3633.709. [DOI] [PubMed] [Google Scholar]
  14. Lowy D. R., Rands E., Scolnick E. M. Helper-independent transformation by unintegrated Harvey sarcoma virus DNA. J Virol. 1978 May;26(2):291–298. doi: 10.1128/jvi.26.2.291-298.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mandel M., Higa A. Calcium-dependent bacteriophage DNA infection. J Mol Biol. 1970 Oct 14;53(1):159–162. doi: 10.1016/0022-2836(70)90051-3. [DOI] [PubMed] [Google Scholar]
  16. Miller J. H. Mutational specificity in bacteria. Annu Rev Genet. 1983;17:215–238. doi: 10.1146/annurev.ge.17.120183.001243. [DOI] [PubMed] [Google Scholar]
  17. Mulligan R. C., Berg P. Expression of a bacterial gene in mammalian cells. Science. 1980 Sep 19;209(4463):1422–1427. doi: 10.1126/science.6251549. [DOI] [PubMed] [Google Scholar]
  18. Pellicer A., Robins D., Wold B., Sweet R., Jackson J., Lowy I., Roberts J. M., Sim G. K., Silverstein S., Axel R. Altering genotype and phenotype by DNA-mediated gene transfer. Science. 1980 Sep 19;209(4463):1414–1422. doi: 10.1126/science.7414320. [DOI] [PubMed] [Google Scholar]
  19. Razzaque A., Chakrabarti S., Joffee S., Seidman M. Mutagenesis of a shuttle vector plasmid in mammalian cells. Mol Cell Biol. 1984 Mar;4(3):435–441. doi: 10.1128/mcb.4.3.435. [DOI] [PMC free article] [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. 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]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES