Abstract
We sought an efficient means to introduce specific human chromosomes into stable interspecific hybrid cells for applications in gene mapping and studies of gene regulation. A defective amphotropic retrovirus was used to insert the gene conferring G418 resistance (neo), a dominant selectable marker, into the chromosomes of diploid human fibroblasts, and the marked chromosomes were transferred to mouse recipient cells by microcell fusion. We recovered five microcell hybrid clones containing one or two intact human chromosomes which were identified by karyotype and marker analysis. Integration of the neo gene into a specific human chromosome in four hybrid clones was confirmed by segregation analysis or by in situ hybridization. We recovered four different human chromosomes into which the G418 resistance gene had integrated: human chromosomes 11, 14, 20, and 21. The high efficiency of retroviral vector transformation makes it possible to insert selectable markers into any mammalian chromosomes of interest.
Full text
PDF






Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Athwal R. S., Smarsh M., Searle B. M., Deo S. S. Integration of a dominant selectable marker into human chromosomes and transfer of marked chromosomes to mouse cells by microcell fusion. Somat Cell Mol Genet. 1985 Mar;11(2):177–187. doi: 10.1007/BF01534706. [DOI] [PubMed] [Google Scholar]
- Beaudet A. L., Su T. S., O'Brien W. E., D'Eustachio P., Barker P. E., Ruddle F. H. Dispersion of argininosuccinate-synthetase-like human genes to multiple autosomes and the X chromosome. Cell. 1982 Aug;30(1):287–293. doi: 10.1016/0092-8674(82)90034-4. [DOI] [PubMed] [Google Scholar]
- Cepko C. L., Roberts B. E., Mulligan R. C. Construction and applications of a highly transmissible murine retrovirus shuttle vector. Cell. 1984 Jul;37(3):1053–1062. doi: 10.1016/0092-8674(84)90440-9. [DOI] [PubMed] [Google Scholar]
- 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]
- Cone R. D., Mulligan R. C. High-efficiency gene transfer into mammalian cells: generation of helper-free recombinant retrovirus with broad mammalian host range. Proc Natl Acad Sci U S A. 1984 Oct;81(20):6349–6353. doi: 10.1073/pnas.81.20.6349. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cooper D. N., Smith B. A., Cooke H. J., Niemann S., Schmidtke J. An estimate of unique DNA sequence heterozygosity in the human genome. Hum Genet. 1985;69(3):201–205. doi: 10.1007/BF00293024. [DOI] [PubMed] [Google Scholar]
- Cox D. R., Gedde-Dahl T., Jr Report of the Committee on the Genetic Constitution of Chromosomes 13, 14, 15 and 16. Cytogenet Cell Genet. 1985;40(1-4):206–241. doi: 10.1159/000132175. [DOI] [PubMed] [Google Scholar]
- Donlon T. A., Litt M., Newcom S. R., Magenis R. E. Localization of the restriction fragment length polymorphism D14S1 (pAW-101) to chromosome 14q32.1 leads to 32.2 by in situ hybridization. Am J Hum Genet. 1983 Nov;35(6):1097–1106. [PMC free article] [PubMed] [Google Scholar]
- Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
- Fournier R. E. A general high-efficiency procedure for production of microcell hybrids. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6349–6353. doi: 10.1073/pnas.78.10.6349. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fournier R. E., Frelinger J. A. Construction of microcell hybrid clones containing specific mouse chromosomes: application to autosomes 8 and 17. Mol Cell Biol. 1982 May;2(5):526–534. doi: 10.1128/mcb.2.5.526. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Gilliam T. C., Scambler P., Robbins T., Ingle C., Williamson R., Davies K. E. The positions of three restriction fragment length polymorphisms on chromosome 4 relative to known genetic markers. Hum Genet. 1984;68(2):154–158. doi: 10.1007/BF00279306. [DOI] [PubMed] [Google Scholar]
- Goddard J. M., Caput D., Williams S. R., Martin D. W., Jr Cloning of human purine-nucleoside phosphorylase cDNA sequences by complementation in Escherichia coli. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4281–4285. doi: 10.1073/pnas.80.14.4281. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gusella J. F., Jones C., Kao F. T., Housman D., Puck T. T. Genetic fine-structure mapping in human chromosome 11 by use of repetitive DNA sequences. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7804–7808. doi: 10.1073/pnas.79.24.7804. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gusella J. F., Wexler N. S., Conneally P. M., Naylor S. L., Anderson M. A., Tanzi R. E., Watkins P. C., Ottina K., Wallace M. R., Sakaguchi A. Y. A polymorphic DNA marker genetically linked to Huntington's disease. Nature. 1983 Nov 17;306(5940):234–238. doi: 10.1038/306234a0. [DOI] [PubMed] [Google Scholar]
- 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]
- Killary A. M., Fournier R. E. A genetic analysis of extinction: trans-dominant loci regulate expression of liver-specific traits in hepatoma hybrid cells. Cell. 1984 Sep;38(2):523–534. doi: 10.1016/0092-8674(84)90507-5. [DOI] [PubMed] [Google Scholar]
- 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]
- Nelson D. L., Weis J. H., Przyborski M. J., Mulligan R. C., Seidman J. G., Housman D. E. Metaphase chromosome transfer of introduced selectable markers. J Mol Appl Genet. 1984;2(6):563–577. [PubMed] [Google Scholar]
- Owerbach D., Bell G. I., Rutter W. J., Shows T. B. The insulin gene is located on chromosome 11 in humans. Nature. 1980 Jul 3;286(5768):82–84. doi: 10.1038/286082a0. [DOI] [PubMed] [Google Scholar]
- Rubin C. M., Houck C. M., Deininger P. L., Friedmann T., Schmid C. W. Partial nucleotide sequence of the 300-nucleotide interspersed repeated human DNA sequences. Nature. 1980 Mar 27;284(5754):372–374. doi: 10.1038/284372a0. [DOI] [PubMed] [Google Scholar]
- Ruddle F. H., Creagan R. P. Parasexual approaches to the genetics of man. Annu Rev Genet. 1975;9:407–486. doi: 10.1146/annurev.ge.09.120175.002203. [DOI] [PubMed] [Google Scholar]
- Saxon P. J., Srivatsan E. S., Leipzig G. V., Sameshima J. H., Stanbridge E. J. Selective transfer of individual human chromosomes to recipient cells. Mol Cell Biol. 1985 Jan;5(1):140–146. doi: 10.1128/mcb.5.1.140. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Su T. S., Bock H. G., O'Brien W. E., Beaudet A. L. Cloning of cDNA for argininosuccinate synthetase mRNA and study of enzyme overproduction in a human cell line. J Biol Chem. 1981 Nov 25;256(22):11826–11831. [PubMed] [Google Scholar]
- 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]
- Weis J. H., Nelson D. L., Przyborski M. J., Chaplin D. D., Mulligan R. C., Housman D. E., Seidman J. G. Eukaryotic chromosome transfer: linkage of the murine major histocompatibility complex to an inserted dominant selectable marker. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4879–4883. doi: 10.1073/pnas.81.15.4879. [DOI] [PMC free article] [PubMed] [Google Scholar]
- White R., Leppert M., Bishop D. T., Barker D., Berkowitz J., Brown C., Callahan P., Holm T., Jerominski L. Construction of linkage maps with DNA markers for human chromosomes. Nature. 1985 Jan 10;313(5998):101–105. doi: 10.1038/313101a0. [DOI] [PubMed] [Google Scholar]
- Wijnen L. M., Grzeschik K. H., Pearson P. L., Meera Khan P. The human PGM-2 and its chromosomal localization in man-mouse hybrids. Hum Genet. 1977 Jul 26;37(3):271–278. doi: 10.1007/BF00393608. [DOI] [PubMed] [Google Scholar]
- Wyman A. R., White R. A highly polymorphic locus in human DNA. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6754–6758. doi: 10.1073/pnas.77.11.6754. [DOI] [PMC free article] [PubMed] [Google Scholar]