Skip to main content
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1984 Aug;4(8):1641–1643. doi: 10.1128/mcb.4.8.1641

Short-term, high-efficiency expression of transfected DNA.

D J Sussman, G Milman
PMCID: PMC368960  PMID: 6092925

Abstract

We have achieved high-efficiency uptake and expression of foreign DNA in mouse Ltk- cells by modifying the DEAE-dextran-mediated transfection method of McCutchan and Pagano (J. Natl. Cancer Inst. 42:351-357, 1968) to include an initial incubation at elevated pH followed by a shock treatment with dimethyl sulfoxide. Up to 80% of mouse Ltk- cells transfected with the herpes simplex virus thymidine kinase gene expressed thymidine kinase as measured by autoradiography.

Full text

PDF
1643

Images in this article

Selected References

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

  1. Chu G., Sharp P. A. SV40 DNA transfection of cells in suspension: analysis of efficiency of transcription and translation of T-antigen. Gene. 1981 Mar;13(2):197–202. doi: 10.1016/0378-1119(81)90008-1. [DOI] [PubMed] [Google Scholar]
  2. Gorman C. M., Merlino G. T., Willingham M. C., Pastan I., Howard B. H. The Rous sarcoma virus long terminal repeat is a strong promoter when introduced into a variety of eukaryotic cells by DNA-mediated transfection. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6777–6781. doi: 10.1073/pnas.79.22.6777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Lewis W. H., Srinivasan P. R., Stokoe N., Siminovitch L. Parameters governing the transfer of the genes for thymidine kinase and dihydrofolate reductase into mouse cells using metaphase chromosomes or DNA. Somatic Cell Genet. 1980 May;6(3):333–347. doi: 10.1007/BF01542787. [DOI] [PubMed] [Google Scholar]
  5. McCutchan J. H., Pagano J. S. Enchancement of the infectivity of simian virus 40 deoxyribonucleic acid with diethylaminoethyl-dextran. J Natl Cancer Inst. 1968 Aug;41(2):351–357. [PubMed] [Google Scholar]
  6. Miller C. L., Ruddle F. H. Co-transfer of human X-linked markers into murine somatic cells via isolated metaphase chromosomes. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3346–3350. doi: 10.1073/pnas.75.7.3346. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Milman G., Herzberg M. Efficient DNA transfection and rapid assay for thymidine kinase activity and viral antigenic determinants. Somatic Cell Genet. 1981 Mar;7(2):161–170. doi: 10.1007/BF01567655. [DOI] [PubMed] [Google Scholar]
  8. Parker B. A., Stark G. R. Regulation of simian virus 40 transcription: sensitive analysis of the RNA species present early in infections by virus or viral DNA. J Virol. 1979 Aug;31(2):360–369. doi: 10.1128/jvi.31.2.360-369.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Shulman M., Wilde C. D., Köhler G. A better cell line for making hybridomas secreting specific antibodies. Nature. 1978 Nov 16;276(5685):269–270. doi: 10.1038/276269a0. [DOI] [PubMed] [Google Scholar]
  10. Sompayrac L. M., Danna K. J. Efficient infection of monkey cells with DNA of simian virus 40. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7575–7578. doi: 10.1073/pnas.78.12.7575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Yamaizumi M., Horwich A. L., Ruddle F. H. Expression and stabilization of microinjected plasmids containing the herpes simplex virus thymidine kinase gene and polyoma virus DNA in mouse cells. Mol Cell Biol. 1983 Apr;3(4):511–522. doi: 10.1128/mcb.3.4.511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Zetterberg A., Engström W. Mitogenic effect of alkaline pH on quiescent, serum-starved cells. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4334–4338. doi: 10.1073/pnas.78.7.4334. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES