Abstract
Eighteen variants recovered from specific locus mutation rate experiments involving the mutagen chlorambucil were subjected to several genetic and molecular analyses. Most mutations were found to be homozygous lethal. Because lethality is often presumptive evidence for multilocus-deletion events, 10 mutations were analyzed by Southern blot analysis with probes at, or closely linked to, several of the specific locus test markers, namely, albino (c), brown (b), and dilute (d). All eight mutations (two c; three b; two d; and one dilute-short ear [Df(d se)]) that arose in post-spermatogonial germ cells were deleted for DNA sequences. No evidence for deletion of two d-se region probes was obtained for the remaining two d mutations that arose in stem-cell spermatogonia. Six of the primary mutants also produced low litter sizes ("semisterility"). Karyotypic analysis has, to date, confirmed the presence of reciprocal translocations in four of the six. The high frequency of deletions and translocations among the mutations induced in post-spermatogonial stages by chlorambucil, combined with its overall high efficiency in inducing mutations in these stages, should make chlorambucil mutagenesis useful for generating experimentally valuable germ-line deletions throughout the mouse genome.
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- Copeland N. G., Hutchison K. W., Jenkins N. A. Excision of the DBA ecotropic provirus in dilute coat-color revertants of mice occurs by homologous recombination involving the viral LTRs. Cell. 1983 Jun;33(2):379–387. doi: 10.1016/0092-8674(83)90419-1. [DOI] [PubMed] [Google Scholar]
- Dryja T. P., Rapaport J. M., Joyce J. M., Petersen R. A. Molecular detection of deletions involving band q14 of chromosome 13 in retinoblastomas. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7391–7394. doi: 10.1073/pnas.83.19.7391. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Eckert K. A., Ingle C. A., Klinedinst D. K., Drinkwater N. R. Molecular analysis of mutations induced in human cells by N-ethyl-N-nitrosourea. Mol Carcinog. 1988;1(1):50–56. doi: 10.1002/mc.2940010111. [DOI] [PubMed] [Google Scholar]
- Friend S. H., Bernards R., Rogelj S., Weinberg R. A., Rapaport J. M., Albert D. M., Dryja T. P. A human DNA segment with properties of the gene that predisposes to retinoblastoma and osteosarcoma. Nature. 1986 Oct 16;323(6089):643–646. doi: 10.1038/323643a0. [DOI] [PubMed] [Google Scholar]
- Geissler E. N., Cheng S. V., Gusella J. F., Housman D. E. Genetic analysis of the dominant white-spotting (W) region on mouse chromosome 5: identification of cloned DNA markers near W. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9635–9639. doi: 10.1073/pnas.85.24.9635. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Generoso W. M., Bishop J. B., Gosslee D. G., Newell G. W., Sheu C. J., von Halle E. Heritable translocation test in mice. Mutat Res. 1980 Sep;76(2):191–215. doi: 10.1016/0165-1110(80)90010-x. [DOI] [PubMed] [Google Scholar]
- Jackson I. J. A cDNA encoding tyrosinase-related protein maps to the brown locus in mouse. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4392–4396. doi: 10.1073/pnas.85.12.4392. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jenkins N. A., Copeland N. G., Taylor B. A., Lee B. K. Dilute (d) coat colour mutation of DBA/2J mice is associated with the site of integration of an ecotropic MuLV genome. Nature. 1981 Oct 1;293(5831):370–374. doi: 10.1038/293370a0. [DOI] [PubMed] [Google Scholar]
- Jenkins N. A., Copeland N. G., Taylor B. A., Lee B. K. Organization, distribution, and stability of endogenous ecotropic murine leukemia virus DNA sequences in chromosomes of Mus musculus. J Virol. 1982 Jul;43(1):26–36. doi: 10.1128/jvi.43.1.26-36.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Johnson D. K., Hand R. E., Jr, Rinchik E. M. Molecular mapping within the mouse albino-deletion complex. Proc Natl Acad Sci U S A. 1989 Nov;86(22):8862–8866. doi: 10.1073/pnas.86.22.8862. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Koenig M., Hoffman E. P., Bertelson C. J., Monaco A. P., Feener C., Kunkel L. M. Complete cloning of the Duchenne muscular dystrophy (DMD) cDNA and preliminary genomic organization of the DMD gene in normal and affected individuals. Cell. 1987 Jul 31;50(3):509–517. doi: 10.1016/0092-8674(87)90504-6. [DOI] [PubMed] [Google Scholar]
- Kunkel L. M., Monaco A. P., Middlesworth W., Ochs H. D., Latt S. A. Specific cloning of DNA fragments absent from the DNA of a male patient with an X chromosome deletion. Proc Natl Acad Sci U S A. 1985 Jul;82(14):4778–4782. doi: 10.1073/pnas.82.14.4778. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kwon B. S., Wakulchik M., Haq A. K., Halaban R., Kestler D. Sequence analysis of mouse tyrosinase cDNA and the effect of melanotropin on its gene expression. Biochem Biophys Res Commun. 1988 Jun 30;153(3):1301–1309. doi: 10.1016/s0006-291x(88)81370-6. [DOI] [PubMed] [Google Scholar]
- Lyon M. F., Glenister P. H., Loutit J. F., Evans E. P., Peters J. A presumed deletion covering the W and Ph loci of the mouse. Genet Res. 1984 Oct;44(2):161–168. doi: 10.1017/s0016672300026367. [DOI] [PubMed] [Google Scholar]
- Monaco A. P., Bertelson C. J., Middlesworth W., Colletti C. A., Aldridge J., Fischbeck K. H., Bartlett R., Pericak-Vance M. A., Roses A. D., Kunkel L. M. Detection of deletions spanning the Duchenne muscular dystrophy locus using a tightly linked DNA segment. 1985 Aug 29-Sep 4Nature. 316(6031):842–845. doi: 10.1038/316842a0. [DOI] [PubMed] [Google Scholar]
- Niswander L., Yee D., Rinchik E. M., Russell L. B., Magnuson T. The albino-deletion complex in the mouse defines genes necessary for development of embryonic and extraembryonic ectoderm. Development. 1989 Jan;105(1):175–182. doi: 10.1242/dev.105.1.175. [DOI] [PubMed] [Google Scholar]
- Page D. C., Mosher R., Simpson E. M., Fisher E. M., Mardon G., Pollack J., McGillivray B., de la Chapelle A., Brown L. G. The sex-determining region of the human Y chromosome encodes a finger protein. Cell. 1987 Dec 24;51(6):1091–1104. doi: 10.1016/0092-8674(87)90595-2. [DOI] [PubMed] [Google Scholar]
- Pathak S., Stock A. D., Lusby A. A combination of sister chromatid differential staining and giemsa banding. Experientia. 1975 Aug 15;31(8):916–918. doi: 10.1007/BF02358850. [DOI] [PubMed] [Google Scholar]
- Popp R. A., Bailiff E. G., Skow L. C., Johnson F. M., Lewis S. E. Analysis of a mouse alpha-globin gene mutation induced by ethylnitrosourea. Genetics. 1983 Sep;105(1):157–167. doi: 10.1093/genetics/105.1.157. [DOI] [PMC free article] [PubMed] [Google Scholar]
- RUSSELL W. L. X-ray-induced mutations in mice. Cold Spring Harb Symp Quant Biol. 1951;16:327–336. doi: 10.1101/sqb.1951.016.01.024. [DOI] [PubMed] [Google Scholar]
- Rinchik E. M., Carpenter D. A., Selby P. B. A strategy for fine-structure functional analysis of a 6- to 11-centimorgan region of mouse chromosome 7 by high-efficiency mutagenesis. Proc Natl Acad Sci U S A. 1990 Feb;87(3):896–900. doi: 10.1073/pnas.87.3.896. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rinchik E. M., Machanoff R., Cummings C. C., Johnson D. K. Molecular cloning and mapping of the ecotropic leukemia provirus Emv-23 provides molecular access to the albino-deletion complex in mouse chromosome 7. Genomics. 1989 Apr;4(3):251–258. doi: 10.1016/0888-7543(89)90328-5. [DOI] [PubMed] [Google Scholar]
- Rinchik E. M., Russell L. B., Copeland N. G., Jenkins N. A. Molecular genetic analysis of the dilute-short ear (d-se) region of the mouse. Genetics. 1986 Feb;112(2):321–342. doi: 10.1093/genetics/112.2.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Royer-Pokora B., Kunkel L. M., Monaco A. P., Goff S. C., Newburger P. E., Baehner R. L., Cole F. S., Curnutte J. T., Orkin S. H. Cloning the gene for an inherited human disorder--chronic granulomatous disease--on the basis of its chromosomal location. Nature. 1986 Jul 3;322(6074):32–38. doi: 10.1038/322032a0. [DOI] [PubMed] [Google Scholar]
- Ruppert S., Müller G., Kwon B., Schütz G. Multiple transcripts of the mouse tyrosinase gene are generated by alternative splicing. EMBO J. 1988 Sep;7(9):2715–2722. doi: 10.1002/j.1460-2075.1988.tb03125.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Russell L. B. Definition of functional units in a small chromosomal segment of the mouse and its use in interpreting the nature of radiation-induced mutations. Mutat Res. 1971 Jan;11(1):107–123. doi: 10.1016/0027-5107(71)90036-4. [DOI] [PubMed] [Google Scholar]
- Russell L. B. Functional and structural analyses of mouse genomic regions screened by the morphological specific-locus test. Mutat Res. 1989 May;212(1):23–32. doi: 10.1016/0027-5107(89)90019-5. [DOI] [PubMed] [Google Scholar]
- Russell L. B., Hunsicker P. R., Cacheiro N. L., Bangham J. W., Russell W. L., Shelby M. D. Chlorambucil effectively induces deletion mutations in mouse germ cells. Proc Natl Acad Sci U S A. 1989 May;86(10):3704–3708. doi: 10.1073/pnas.86.10.3704. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Russell L. B., Montgomery C. S., Raymer G. D. Analysis of the albino-locus region of the mouse: IV. Characterization of 34 deficiencies. Genetics. 1982 Mar;100(3):427–453. doi: 10.1093/genetics/100.3.427. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Russell W. L., Kelly E. M., Hunsicker P. R., Bangham J. W., Maddux S. C., Phipps E. L. Specific-locus test shows ethylnitrosourea to be the most potent mutagen in the mouse. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5818–5819. doi: 10.1073/pnas.76.11.5818. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sega G. A., Sotomayor R. E., Owens J. G. A study of unscheduled DNA synthesis induced by X-rays in the germ cells of male mice. Mutat Res. 1978 Feb;49(2):239–257. doi: 10.1016/0027-5107(78)90163-x. [DOI] [PubMed] [Google Scholar]
- Shibahara S., Tomita Y., Sakakura T., Nager C., Chaudhuri B., Müller R. Cloning and expression of cDNA encoding mouse tyrosinase. Nucleic Acids Res. 1986 Mar 25;14(6):2413–2427. doi: 10.1093/nar/14.6.2413. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Siracusa L. D., Russell L. B., Jenkins N. A., Copeland N. G. Allelic variation within the Emv-15 locus defines genomic sequences closely linked to the agouti locus on mouse chromosome 2. Genetics. 1987 Sep;117(1):85–92. doi: 10.1093/genetics/117.1.85. [DOI] [PMC free article] [PubMed] [Google Scholar]