Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1985 Jan;5(1):140–146. doi: 10.1128/mcb.5.1.140

Selective transfer of individual human chromosomes to recipient cells.

P J Saxon, E S Srivatsan, G V Leipzig, J H Sameshima, E J Stanbridge
PMCID: PMC366688  PMID: 3982414

Abstract

Two hypoxanthine phosphoribosyltransferase-deficient human cell lines, D98/AH-2 and HT1080-6TG, were stably transfected with pSV2 gpt, a plasmid containing the selectable marker Escherichia coli xanthine-guanine phosphoribosyl transferase (Eco gpt). Hypoxanthine-aminopterin-thymidine-resistant transformants arose with a frequency of ca. 10(-6) and contained mostly single, but occasionally multiple, copies of the plasmid sequences. These transformants actively express the Eco gpt marker. Single chromosomes from two different HT1080 gpt transformants and one D98 gpt transformant, containing the integrated plasmid sequences, were transferred via microcell-mediated chromosome transfer to hypoxanthine phosphoribosyl transferase-deficient mouse A9 cells. The transferred human chromosomes were identified as 2, 4, and 22, by using a combination of G-11 staining, G-banding, isoenzyme analysis, and in situ hybridization. This system is being used to create a library of interspecies microcell hybrid clones, each clone containing a unique single human chromosome in a mouse background. The complete library will represent the entire human karyotype.

Full text

PDF
140

Images in this article

142Image
on p.142
142Image
on p.142
143Image
on p.143
143Image
on p.143
144Image
on p.144
145Image
on p.145

Selected References

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

  1. Botstein D., White R. L., Skolnick M., Davis R. W. Construction of a genetic linkage map in man using restriction fragment length polymorphisms. Am J Hum Genet. 1980 May;32(3):314–331. [PMC free article] [PubMed] [Google Scholar]
  2. Chasin L. A., Urlaub G. Mutant alleles for hypoxanthine phosphoriboxyltransferase: codominant expression, complementation, and segregation in hybrid Chinese hamster cells. Somatic Cell Genet. 1976 Sep;2(5):453–467. doi: 10.1007/BF01542725. [DOI] [PubMed] [Google Scholar]
  3. Cirullo R. E., Dana S., Wasmuth J. J. Efficient procedure for transferring specific human genes into Chinese hamster cell mutants: interspecific transfer of the human genes encoding leucyl- and asparaginyl-tRNA synthetases. Mol Cell Biol. 1983 May;3(5):892–902. doi: 10.1128/mcb.3.5.892. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Creagan R. P., Ruddle F. H. The clone panel: a systematic approach to gene mapping using interspecific somatic cell hybrids. Cytogenet Cell Genet. 1975;14(3-6):282–286. doi: 10.1159/000130364. [DOI] [PubMed] [Google Scholar]
  5. Croce C. M. Loss of mouse chromosomes in somatic cell hybrids between HT-1080 human fibrosarcoma cells and mouse peritioneal macrophages. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3248–3252. doi: 10.1073/pnas.73.9.3248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ege T., Ringertz N. R., Hamberg H., Sidebottom E. Preparation of microcells. Methods Cell Biol. 1977;15:339–357. [PubMed] [Google Scholar]
  7. Fournier R. E., Ruddle F. H. Microcell-mediated transfer of murine chromosomes into mouse, Chinese hamster, and human somatic cells. Proc Natl Acad Sci U S A. 1977 Jan;74(1):319–323. doi: 10.1073/pnas.74.1.319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Friend K. K., Chen S., Ruddle F. H. Differential staining of interspecific chromosomes in somatic cell hybrids by alkaline Giemsa stain. Somatic Cell Genet. 1976 Mar;2(2):183–188. doi: 10.1007/BF01542631. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Harper M. E., Saunders G. F. Localization of single copy DNA sequences of G-banded human chromosomes by in situ hybridization. Chromosoma. 1981;83(3):431–439. doi: 10.1007/BF00327364. [DOI] [PubMed] [Google Scholar]
  11. Irwin M., Oates D. C., Patterson D. Biochemical genetics of Chinese hamster cell mutants with deviant purine metabolism: isolation and characterization of a mutant deficient in the activity of phosphoribosylaminoimidazole synthetase. Somatic Cell Genet. 1979 Mar;5(2):203–216. doi: 10.1007/BF01539161. [DOI] [PubMed] [Google Scholar]
  12. Jones C., Patterson D., Kao F. T. Assignment of the gene coding for phosphoribosylglycineamide formyltransferase to human chromosome 14. Somatic Cell Genet. 1981 Jul;7(4):399–409. doi: 10.1007/BF01542985. [DOI] [PubMed] [Google Scholar]
  13. Kao F. T., Jones C., Puck T. T. Genetics of somatic mammalian cells: genetic, immunologic, and biochemical analysis with Chinese hamster cell hybrids containing selected human chromosomes. Proc Natl Acad Sci U S A. 1976 Jan;73(1):193–197. doi: 10.1073/pnas.73.1.193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Littlefield J. W. The use of drug-resistant markers to study the hybridization of mouse fibroblasts. Exp Cell Res. 1966 Jan;41(1):190–196. doi: 10.1016/0014-4827(66)90558-1. [DOI] [PubMed] [Google Scholar]
  16. McBride O. W., Ozer H. L. Transfer of genetic information by purified metaphase chromosomes. Proc Natl Acad Sci U S A. 1973 Apr;70(4):1258–1262. doi: 10.1073/pnas.70.4.1258. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. McNeill C. A., Brown R. L. Genetic manipulation by means of microcell-mediated transfer of normal human chromosomes into recipient mouse cells. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5394–5398. doi: 10.1073/pnas.77.9.5394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Rasheed S., Nelson-Rees W. A., Toth E. M., Arnstein P., Gardner M. B. Characterization of a newly derived human sarcoma cell line (HT-1080). Cancer. 1974 Apr;33(4):1027–1033. doi: 10.1002/1097-0142(197404)33:4<1027::aid-cncr2820330419>3.0.co;2-z. [DOI] [PubMed] [Google Scholar]
  20. Ruddle F. H. A new era in mammalian gene mapping: somatic cell genetics and recombinant DNA methodologies. Nature. 1981 Nov 12;294(5837):115–120. doi: 10.1038/294115a0. [DOI] [PubMed] [Google Scholar]
  21. Russell W. C., Newman C., Williamson D. H. A simple cytochemical technique for demonstration of DNA in cells infected with mycoplasmas and viruses. Nature. 1975 Feb 6;253(5491):461–462. doi: 10.1038/253461a0. [DOI] [PubMed] [Google Scholar]
  22. Sandri-Goldin R. M., Goldin A. L., Levine M., Glorioso J. C. High-frequency transfer of cloned herpes simplex virus type 1 sequences to mammalian cells by protoplast fusion. Mol Cell Biol. 1981 Aug;1(8):743–752. doi: 10.1128/mcb.1.8.743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schwab M., Varmus H. E., Bishop J. M., Grzeschik K. H., Naylor S. L., Sakaguchi A. Y., Brodeur G., Trent J. Chromosome localization in normal human cells and neuroblastomas of a gene related to c-myc. Nature. 1984 Mar 15;308(5956):288–291. doi: 10.1038/308288a0. [DOI] [PubMed] [Google Scholar]
  24. Sekiguchi T., Sekiguchi F., Tomii S. Genetic complementation in hybrid cells derived from mutagen-induced mouse clones deficient in HGPRT activity. Exp Cell Res. 1975 Jun;93(1):207–218. doi: 10.1016/0014-4827(75)90441-3. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Srivatsan E. S., Stanbridge E. J., Saxon P. J., Stambrook P. J., Trill J. J., Tischfield J. A. Plasmid, phage, and genomic DNA-mediated transfer and expression of prokaryotic and eukaryotic genes in cultured human cells. Cytogenet Cell Genet. 1984;38(3):227–234. doi: 10.1159/000132065. [DOI] [PubMed] [Google Scholar]
  27. Tunnacliffe A., Parkar M., Povey S., Bengtsson B. O., Stanley K., Solomon E., Goodfellow P. Integration of Ecogpt and SV40 early region sequences into human chromosome 17: a dominant selection system in whole cell and microcell human-mouse hybrids. EMBO J. 1983;2(9):1577–1584. doi: 10.1002/j.1460-2075.1983.tb01627.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Watson B., Gormley I. P., Gardiner S. E., Evans H. J., Harris H. Reappearance of murine hypoxanthine guanine phosphoribosyl transferase activity in mouse A9 cells after attempted hybridisation with human cell lines. Exp Cell Res. 1972 Dec;75(2):401–409. doi: 10.1016/0014-4827(72)90446-6. [DOI] [PubMed] [Google Scholar]
  29. Weiss M. C., Green H. Human-mouse hybrid cell lines containing partial complements of human chromosomes and functioning human genes. Proc Natl Acad Sci U S A. 1967 Sep;58(3):1104–1111. doi: 10.1073/pnas.58.3.1104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Worton R. G., Duff C. Karyotyping. Methods Enzymol. 1979;58:322–344. doi: 10.1016/s0076-6879(79)58148-8. [DOI] [PubMed] [Google Scholar]
  31. Yu R. L., Aronson M. M., Nichols W. W. High-resolution bands in human fibroblast chromosomes induced by actinomycin D. Cytogenet Cell Genet. 1981;31(2):111–114. doi: 10.1159/000131634. [DOI] [PubMed] [Google Scholar]

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

RESOURCES