Abstract
Recombinant bacteriophage lambda from a human genomic library were screened to indentify human DNA inserts having only unique sequences. Unique human inserts were found in about 1% of the phage screened. One recombinant phage, P3-2, was studied in detail. It contains a human insert of 14.7 kilobases with four internal EcoRI cleavage sites. A restriction map was constructed for EcoRI and BamHI sites. Hybridization of the 32P-labeled P3-2 probe to a Southern blot of EcoRI-digested total human DNA yielded distinct bands at positions corresponding to the human insert fragments contained in P3-2. By using a series of human-Chinese hamster somatic cell hybrids containing unique combinations of human chromosomes, the human DNA segment in phage P3-2 was assigned to human chromosome 22 by blot hybridization and synteny analysis. In addition, another human DNA segment, 11.4 kilobases, in phage P3-10 was assigned to human chromosome 10 by similar procedures. With this approach, more unique DNA sequences can be isolated, assigned to specific human chromosomes, and used as genetic markers for gene mapping and linkage, polymorphism, and other genetic studies in the human genome.
Full text
PDF![865](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/618d/345853/ec55313a2a14/pnas00442-0153.png)
![866](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/618d/345853/75a9e2e954bf/pnas00442-0154.png)
![867](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/618d/345853/fdc59b3eab3f/pnas00442-0155.png)
![868](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/618d/345853/238691a6e2ff/pnas00442-0156.png)
![869](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/618d/345853/464eb14c07c2/pnas00442-0157.png)
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Danna K. J., Sack G. H., Jr, Nathans D. Studies of simian virus 40 DNA. VII. A cleavage map of the SV40 genome. J Mol Biol. 1973 Aug 5;78(2):363–376. doi: 10.1016/0022-2836(73)90122-8. [DOI] [PubMed] [Google Scholar]
- Gusella J. F., Keys C., VarsanyiBreiner A., Kao F. T., Jones C., Puck T. T., Housman D. Isolation and localization of DNA segments from specific human chromosomes. Proc Natl Acad Sci U S A. 1980 May;77(5):2829–2833. doi: 10.1073/pnas.77.5.2829. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gusella J., Varsanyi-Breiner A., Kao F. T., Jones C., Puck T. T., Keys C., Orkin S., Housman D. Precise localization of human beta-globin gene complex on chromosome 11. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5239–5242. doi: 10.1073/pnas.76.10.5239. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Harper M. E., Ullrich A., Saunders G. F. Localization of the human insulin gene to the distal end of the short arm of chromosome 11. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4458–4460. doi: 10.1073/pnas.78.7.4458. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jones C., Kao F. T., Taylor R. T. Chromosomal assignment of the gene for folylpolyglutamate synthetase to human chromosome 9. Cytogenet Cell Genet. 1980;28(3):181–194. doi: 10.1159/000131529. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Kao F. T., Puck T. T. Genetics of somatic mammalian cells. XIV. Genetic analysis in vitro of auxotrophic mutants. J Cell Physiol. 1972 Aug;80(1):41–50. doi: 10.1002/jcp.1040800106. [DOI] [PubMed] [Google Scholar]
- Law M. L., Kao F. T. Induced segregation of human syntenic genes by 5-bromodeozyuridine + near-visible light. Somatic Cell Genet. 1978 Jul;4(4):465–476. doi: 10.1007/BF01538867. [DOI] [PubMed] [Google Scholar]
- Lawn R. M., Fritsch E. F., Parker R. C., Blake G., Maniatis T. The isolation and characterization of linked delta- and beta-globin genes from a cloned library of human DNA. Cell. 1978 Dec;15(4):1157–1174. doi: 10.1016/0092-8674(78)90043-0. [DOI] [PubMed] [Google Scholar]
- Lebo R. V., Carrano A. V., Burkhart-Schultz K., Dozy A. M., Yu L. C., Kan Y. W. Assignment of human beta-, gamma-, and delta-globin genes to the short arm of chromosome 11 by chromosome sorting and DNA restriction enzyme analysis. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5804–5808. doi: 10.1073/pnas.76.11.5804. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moore E. E., Jones C., Kao F. T., Oates D. C. Synteny between glycinamide ribonucleotide synthetase and superoxide dismutase (soluble). Am J Hum Genet. 1977 Jul;29(4):389–396. [PMC free article] [PubMed] [Google Scholar]
- Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [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]
- Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [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]