Home LiteratureArticle Details
PMID: 6132383 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Homologous recombination in mammalian cells mediates formation of a functional gene from two overlapping gene fragments.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 80 ·No. 7 ·1983-04-00 ·Pages 2002-6

de Saint Vincent BR, Wahl GM

Abstract

Chinese hamster cells with a lesion in the CAD gene (cell line Urd-A) require exogenous uridine to survive. Uridine prototrophs could be isolated after introducing two recombinant plasmids containing overlapping fragments of a cloned Syrian hamster CAD gene. In contrast, no uridine prototrophs were obtained after introducing a plasmid containing only one of the two overlapping fragments. DNA restriction analysis showed that the prototrophic transformants contain a functional CAD gene which was formed by a recombination event in the overlapping region of the two clones. Most of the recombination events involved homologous exchanges, and some of them apparently were reciprocal. In situ hybridization analysis revealed that the donated sequences were integrated at a single chromosomal site which was different in each transformant. These results demonstrate the existence of a recombination system(s) in mammalian cells that can catalyze homologous exchanges. Recombination between donated sequences is a means by which this system can be characterized and also utilized for the production of novel gene fusions.

MeSH Terms
Animals Aspartate Carbamoyltransferase/genetics Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing)/genetics Cricetinae Crossing Over, Genetic Dihydroorotase/genetics Genes Multienzyme Complexes/genetics Plasmids Proteins/genetics Recombination, Genetic Sister Chromatid Exchange Uridine/biosynthesis
Chemicals
CAD trifunctional enzyme Multienzyme Complexes Proteins Aspartate Carbamoyltransferase Dihydroorotase Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing) Uridine
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
de Saint Vincent B R
Wahl G M
References (33)
33 references, click to expand
  1. Genetic recombination and commitment to meiosis in Saccharomyces.
    Proc Natl Acad Sci U S A. 1974 Aug;71(8):3172-6 PMID: 4606582
  2. Evidence for mitotic recombination in the cellular slime mold Dictyostelium discoideum.
    Proc Natl Acad Sci U S A. 1974 Oct;71(10):4025-6 PMID: 4530283
  3. Enhanced mitotic recombination in a ligase-defective mutant of the yeast Saccharomyces cerevisiae.
    Proc Natl Acad Sci U S A. 1979 Sep;76(9):4589-92 PMID: 388447
  4. Single-copy and amplified CAD genes in Syrian hamster chromosomes localized by a highly sensitive method for in situ hybridization.
    Mol Cell Biol. 1982 Mar;2(3):308-19 PMID: 6180304
  5. ABERRANT RECOMBINATION OF PYRIDOXINE MUTANTS OF Neurospora.
    Proc Natl Acad Sci U S A. 1955 Apr 15;41(4):215-20 PMID: 16589648
  6. Segregation of recessive phenotypes in somatic cell hybrids: role of mitotic recombination, gene inactivation, and chromosome nondisjunction.
    Mol Cell Biol. 1981 Apr;1(4):336-46 PMID: 6965251
  7. DNA sequence homology and chromosomal deletion at a site of SV40 DNA integration.
    Nature. 1982 Mar 25;296(5855):363-6 PMID: 6278325
  8. Somatic Crossing over and Segregation in Drosophila Melanogaster.
    Genetics. 1936 Nov;21(6):625-730 PMID: 17246815
  9. Biochemical controls of meiosis.
    Annu Rev Genet. 1973;7:37-66 PMID: 4593309
  10. CYTOLOGICAL EVIDENCE FOR CROSSING-OVER IN VITRO IN HUMAN LYMPHOID CELLS.
    Science. 1964 Apr 17;144(3616):298-301 PMID: 14169717
  11. Changes in DNA during meiosis in a repair-deficient mutant (rad 52) of yeast.
    Science. 1981 May 1;212(4494):543-5 PMID: 7010606
  12. A subcloning strategy for DNA sequence analysis.
    Nucleic Acids Res. 1980 Dec 11;8(23):5541-9 PMID: 6258146
  13. Properties of dispersed, highly repeated DNA within and near the hamster CAD gene.
    Mol Cell Biol. 1982 Mar;2(3):302-7 PMID: 6180303
  14. Sister Chromatid Exchanges in Tritium-Labeled Chromosomes.
    Genetics. 1958 May;43(3):515-29 PMID: 17247775
  15. Patterns of integration of DNA microinjected into cultured mammalian cells: evidence for homologous recombination between injected plasmid DNA molecules.
    Mol Cell Biol. 1982 Nov;2(11):1372-87 PMID: 6298598
  16. The cloning and reintroduction into animal cells of a functional CAD gene, a dominant amplifiable genetic marker.
    Cell. 1981 Dec;27(2 Pt 1):267-77 PMID: 6277500
  17. Analysis of recombination in mammalian cells using SV40 genome segments having homologous overlapping termini.
    Nucleic Acids Res. 1980 Jun 25;8(12):2725-36 PMID: 6253890
  18. Detection by density equilibrium centrifugation of recombinant-like DNA molecules in somatic mammalian cells.
    J Mol Biol. 1975 Oct 15;98(1):195-218 PMID: 1195379
  19. Evidence for the formation of hybrid DNA during mitotic recombination in Chinese hamster cells.
    Cell. 1976 Aug;8(4):573-9 PMID: 954106
  20. A search for allelic recombination in Chinese hamster cell hybrids.
    Mol Gen Genet. 1977 Nov 18;156(3):273-9 PMID: 600266
  21. Separation of linked markers in Chinese hamster cell hybrids: mitotic recombination is not involved.
    Genetics. 1978 Dec;90(4):735-60 PMID: 744475
  22. Mutations of Bacteria from Virus Sensitivity to Virus Resistance.
    Genetics. 1943 Nov;28(6):491-511 PMID: 17247100
  23. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid.
    J Bacteriol. 1978 Jun;134(3):1141-56 PMID: 149110
  24. Radiation-Induced Genetic Segregations in Vegetative Cells of Diploid Yeast.
    Genetics. 1955 Nov;40(6):826-31 PMID: 17247593
  25. 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):3683-7 PMID: 291033
  26. Structure of the gene for CAD, the multifunctional protein that initiates UMP synthesis in Syrian hamster cells.
    Mol Cell Biol. 1982 Mar;2(3):293-301 PMID: 6125880
  27. Adenovirus recombination in normal and repair-deficient human fibroblasts.
    Virology. 1980 Jan 15;100(1):179-84 PMID: 7350726
  28. A manyfold increase in sister chromatid exchanges in Bloom's syndrome lymphocytes.
    Proc Natl Acad Sci U S A. 1974 Nov;71(11):4508-12 PMID: 4140506
  29. Genetic analysis based on mitotic recombination.
    Adv Genet. 1958;9:71-104 PMID: 13520440
  30. Phenomenology and genetic control of mitotic recombination in yeast.
    Annu Rev Genet. 1981;15:57-89 PMID: 7039496
  31. DNA-mediated gene transfer: recombination between cotransferred DNA sequences and recovery of recombinants in a plasmid.
    Proc Natl Acad Sci U S A. 1982 May;79(9):2748-52 PMID: 6283542
  32. Yeast transformation: a model system for the study of recombination.
    Proc Natl Acad Sci U S A. 1981 Oct;78(10):6354-8 PMID: 6273866
  33. Sister chromatid exchanges, indices of human chromosome damage and repair: detection by fluorescence and induction by mitomycin C.
    Proc Natl Acad Sci U S A. 1974 Aug;71(8):3162-6 PMID: 4137928
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.80.7.2002
Published
1983-04-00
Pages
2002-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC393740
Subset
IM
Grants
NIGMS NIH HHS · GM27754 · United States
External Links
PubMed source DOI: 10.1073/pnas.80.7.2002 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2