Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1975 Sep;72(9):3531–3535. doi: 10.1073/pnas.72.9.3531

Gene transfer to human cells: transducing phage lambda plac gene expression in GMI-gangliosidosis fibroblasts.

J Horst, F Kluge, K Beyreuther, W Gerok
PMCID: PMC433029  PMID: 242006

Abstract

Genetic information from the bacterium Escherichia coli was transferred to human cells by means of the specialized transducing phage lambda plac carrying the bacterial z gene for the enzyme beta-galactosidase (geta-D-galactoside galactohydrolase, EC 3.2.1.23). As recipient cells, cultured skin fibroblasts from a patient with generalized gangliosidosis (GMI-gangliosidosis Type I) characterized by a severe deficiency of beta-galactosidase activity were used. The deficient human cells were incubated with the bacteriophage lambda plac or lambda plac DNA and beta-galactosidase activity was measured in order to detect gene transfer and acceptance of the prokaryotic information in the mammalian system for transcription and translation. The expression of the phage genome in the deficient fibroblasts could be demonstrated by detection of higher beta-galactosidase activity after incubation with phage lambda plac in three out of 19 experiments and in four out of 16 experiments after treatment with lambda plac DNA. Lambda plac DNA induced much higher enzyme activities than infective phage particles. Immunochemical and physicochemical assays could not distinguish the induced beta-galactosidase activity from that of the z-gene product of E. coli.

Full text

PDF
3532

Selected References

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

  1. Anker P., Stroun M. Bacterial ribonucleic acid in the frog brain after a bacterial peritoneal infection. Science. 1972 Nov 10;178(4061):621–623. doi: 10.1126/science.178.4061.621. [DOI] [PubMed] [Google Scholar]
  2. Aviv H., Boime I., Loyd B., Leder P. Translation of bacteriophage Q messenger RNA in a murine Krebs 2 ascites tumor cell-free system. Science. 1972 Dec 22;178(4067):1293–1295. doi: 10.1126/science.178.4067.1293. [DOI] [PubMed] [Google Scholar]
  3. Beyreuther K., Adler K., Geisler N., Klemm A. The amino-acid sequence of lac repressor. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3576–3580. doi: 10.1073/pnas.70.12.3576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Carlson P. S. The use of protoplasts for genetic research. Proc Natl Acad Sci U S A. 1973 Feb;70(2):598–602. doi: 10.1073/pnas.70.2.598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DULBECCO R., VOGT M. One-step growth curve of Western equine encephalomyelitis virus on chicken embryo cells grown in vitro and analysis of virus yields from single cells. J Exp Med. 1954 Feb;99(2):183–199. doi: 10.1084/jem.99.2.183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Doy C. H., Gresshoff P. M., Rolfe B. G. Biological and molecular evidence for the transgenosis of genes from bacteria to plant cells. Proc Natl Acad Sci U S A. 1973 Mar;70(3):723–726. doi: 10.1073/pnas.70.3.723. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Doy C. H., Gresshoff P. M., Rolfe B. G. Time-course of phenotypic expression of Escherichia coli gene Z following transgenosis in haploid Lycopersicon esculentum cells. Nat New Biol. 1973 Jul 18;244(133):90–91. doi: 10.1038/newbio244090a0. [DOI] [PubMed] [Google Scholar]
  8. Geier M. R., Merril C. R. Lambda phage transcription in human fibroblasts. Virology. 1972 Mar;47(3):638–643. doi: 10.1016/0042-6822(72)90553-3. [DOI] [PubMed] [Google Scholar]
  9. Gilbert W., Maxam A. The nucleotide sequence of the lac operator. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3581–3584. doi: 10.1073/pnas.70.12.3581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gilbert W., Müller-Hill B. Isolation of the lac repressor. Proc Natl Acad Sci U S A. 1966 Dec;56(6):1891–1898. doi: 10.1073/pnas.56.6.1891. [DOI] [PMC free article] [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. Groneberg J., Brown D. T., Doerfler W. Uptake and fate of the DNA of adenovirus type 2 in KB cells. Virology. 1975 Mar;64(1):115–131. doi: 10.1016/0042-6822(75)90084-7. [DOI] [PubMed] [Google Scholar]
  13. Hultbery B., Ockerman P. A. Artificial substrates in the assay of acid glycosidases. Clin Chim Acta. 1972 Jun;39(1):49–58. doi: 10.1016/0009-8981(72)90298-7. [DOI] [PubMed] [Google Scholar]
  14. Ippen K., Shapiro J. A., Beckwith J. R. Transposition of the lac region to the gal region of the Escherichia coli chromosome: isolation of lambda-lac transducing bacteriophages. J Bacteriol. 1971 Oct;108(1):5–9. doi: 10.1128/jb.108.1.5-9.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. JACOB F., MONOD J. Genetic regulatory mechanisms in the synthesis of proteins. J Mol Biol. 1961 Jun;3:318–356. doi: 10.1016/s0022-2836(61)80072-7. [DOI] [PubMed] [Google Scholar]
  16. Johnson C. B., Grierson D., Smith H. Expression of lambda plac5 DNA in cultured cells of a higher plant. Nat New Biol. 1973 Jul 25;244(134):105–107. doi: 10.1038/newbio244105a0. [DOI] [PubMed] [Google Scholar]
  17. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  18. Ledoux L., Huart R., Jacobs M. DNA-mediated genetic correction of thiamineless Arabidopsis thaliana. Nature. 1974 May 3;249(452):17–21. doi: 10.1038/249017a0. [DOI] [PubMed] [Google Scholar]
  19. Melchers F., Messer W. Enhanced stability against heat denaturation of E. coli wild type and mutant beta-galactosidase in the presence of specific antibodies. Biochem Biophys Res Commun. 1970 Aug 11;40(3):570–575. doi: 10.1016/0006-291x(70)90940-x. [DOI] [PubMed] [Google Scholar]
  20. Merril C. R., Geier M. R., Petricciani J. C. Bacterial virus gene expression in human cells. Nature. 1971 Oct 8;233(5319):398–400. doi: 10.1038/233398a0. [DOI] [PubMed] [Google Scholar]
  21. Morrison T. G., Lodish H. F. Translation of bacteriophage Q RNA by cytoplasmic extracts of mammalian cells. Proc Natl Acad Sci U S A. 1973 Feb;70(2):315–319. doi: 10.1073/pnas.70.2.315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Norden A. G., O'Brien J. S. Ganglioside GM1 beta-galactosidase: studies in human liver and brain. Arch Biochem Biophys. 1973 Nov;159(1):383–392. doi: 10.1016/0003-9861(73)90465-7. [DOI] [PubMed] [Google Scholar]
  23. Richter D., Herrlich P., Schweiger M. Phage DNA directed enzyme synthesis in vitro system from yeast mitochondria. Nat New Biol. 1972 Jul 19;238(81):74–76. doi: 10.1038/newbio238074a0. [DOI] [PubMed] [Google Scholar]
  24. Robins E., Hirsch H. E., Emmons S. S. Glycosidases in the nervous system. I. Assay, some properties, and distribution of beta-galactosidase, beta-glucoronidase, and beta-glucosidase. J Biol Chem. 1968 Aug 25;243(16):4246–4252. [PubMed] [Google Scholar]
  25. Sander E. Alteration of fd phage in tobacco leaves. Virology. 1967 Sep;33(1):121–130. doi: 10.1016/0042-6822(67)90100-6. [DOI] [PubMed] [Google Scholar]
  26. Schreier M. H., Staehelin T., Gesteland R. F., Spahr P. F. Translation of bacteriophage R17 and Qbeta RNA in a mammalian cell-free system. J Mol Biol. 1973 Apr 15;75(3):575–578. doi: 10.1016/0022-2836(73)90462-2. [DOI] [PubMed] [Google Scholar]
  27. Studzinski G. P., Gierthy J. F., Cholon J. J. An autoradiographic screening test for mycoplasmal contamination of mammalian cell cultures. In Vitro. 1973 May-Jun;8(6):466–472. doi: 10.1007/BF02615948. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES