Abstract
Using degenerate primer amplification of chromosomes microdissected from banded cytogenetic preparations, we constructed both whole chromosome painting probes for mouse Chromosomes (Chrs) 1, 2, 3, and 11 and a centromere probe that strongly paints most mouse centromeres. We also amplified a Robertsonian translocation chromosome microdissected from unstained preparations to construct a painting probe for Chrs 9 and 19. The chromosomes probes uniformly painted the respective chromosomes of origin. We demonstrated the utility of the Chr 11 probe in aberration analysis by staining mutants that we had previously identified as containing a Chr 11 translocation, and in some mutant cell lines we observed chromosome rearrangements not previously detected in stained cytogenetic preparations. The technology of microdissection and amplification applies to all mouse chromosomes or to specific subchromosomal regions and will be useful in mouse genetics, in aberration analysis, and for chromosome identification.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Breneman, J.W., Ramsey, M.J., Lee, D.A., Eveleth, G.G., Minkler, J.L., Tucker, J.D. (1993). The development of chromosome-specific composite DNA probes for the mouse and their application to chromosome painting. Chromosoma 102, 591–598.
Carrano, A.V. (1973). Chromosome aberrations and radiation-induced cell death. I. Transmission and survival parameters of aberrations. Mutat. Res. 17, 341–353.
Cremer, T., Lichter, P., Borden, J., Ward, D.C., Manuelidis, L. (1988). Detection of chromosome aberrations in metaphase and interphase tumor cells by hybridization using chromosome-specific library probes. Hum. Genet. 80, 235–246.
Guan, X.-Y., Trent, J.M., Meltzer, P.S. (1993). Generation of band-specific painting probes from a single microdissected chromosome. Hum. Mol. Genet. 2, 1117–1121.
Guan, X.-Y., Meltzer, P.S., Trent, J. (1994). Rapid generation of whole chromosome painting probes (WCPs) by chromosome microdissection. Genomics 22, 101–107.
Hozier, J.C., Davis, L.M. (1992). Cytogenetic approaches to genome mapping. Anal. Biochem. 200, 205–217.
Hozier, J., Sawyer, J., Moore, M., Howard, B., Clive, D. (1981). Cytogenetic analysis of the L5178Y/TK+/- → TK+/- mouse lymphoma mutagenesis assay system. Mutat. Res. 84, 169–181.
Hozier, J., Sawyer, J., Clive, D., Moore, M. (1982). Cytogenetic distinction between the TK+ and TK- chromosomes in the L5178Y TK+/-3.7.2C mouse-lymphoma cell line. Mutat. Res. 105, 451–456.
Hozier, J., Graham, R., Westfall, T., Siebert, P., Davis, L. (1994). Preparative in situ hybridization: selection of chromosome region-specific libraries on mitotic chromosomes. Genomics 19, 441–447.
Ledbetter, S.A., Nelson, D.L., Warren, S.T., Ledbetter, D.H. (1990). Rapid isolation of DNA probes within specific chromosome regions by interspersed repetitive sequence polymerase chain reaction. Genomics 6, 475–481.
Lucas, J.N., Awa, A., Straume, T., Poggensee, M., Kodama, Y., Nakano, M., Ohtaki, K., Weier, H.-U., Pinkel, D., Gray, J., Littlefield, G. (1992). Rapid translocation frequency analysis in humans decades after exposure to ionizing radiation. Int. J. Radiat. Biol. 62, 53–63.
Meltzer, P.S., Guan, X.-Y., Burgess, A., Trent, J.M. (1992). Rapid generation of region specific probes by chromosome microdissection and their application. Nature Genet. 1, 24–28.
Miyashita, K., Voojis, M.A., Tucker, J.D., Lee, D.A., Gray, J.W., Pallavicini, M.G. (1994). A mouse chromosome 11 library generated from sorted chromosomes using linker-adapter polymerase chain reaction. Cytogenet. Cell Genet. 66, 54–57.
Moore, M.M., Clive, D., Hozier, J.C., Howard, B.E., Batson, A.G., Turner, N.T. Sawyer, J. (1985). Analysis of trifluorothymidine-resistant (TFTr) mutants of L5178Y/TK+/- mouse lymphoma cells. Mutat. Res. 151, 161–174.
Preston, R.J. (1992). Taking advantage of current trends in biology. Environ. Mutagen. Soc. Newslett., September.
Sawyer, J.R., Moore, M.M., Hozier, J.C. (1987). High resolution G-banded chromosomes of the mouse. Chromosoma 95, 350–358.
Telenius, H., Carter, N.P., Bebb, C.E., Nordenskjold, M., Ponder, B.A.J., Tunnacliffe, A. (1992). Degenerate oligonucleotide-primed PCR (DOP-PCR): general amplification of target DNA by a single degenerate primer. Genomics 13, 718–725.
Weier, H.U.G., Polikoff, D., Fawcett, J.J., Greulich, K.M., Lee, K.H., Cram, S., Chapman, V.M., Gray, J.W. (1994). Generation of five high-complexity painting probe libraries from flow-sorted mouse chromosomes. Genomics 21, 641–644.
Zhang, J., Meltzer, P., Jenkins, R., Guan, X.-Y., Trent, J. (1993). Application of chromosome microdissection probes for elucidation of BCR-ABL fusion and variant Philadelphia chromosome translocations in chronic myelogenous leukemia. Blood 81, 3365–3371.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Liechty, M.C., Hall, B.K., Scalzi, J.M. et al. Mouse chromosome-specific painting probes generated from microdissected chromosomes. Mammalian Genome 6, 592–594 (1995). https://doi.org/10.1007/BF00352363
Issue Date:
DOI: https://doi.org/10.1007/BF00352363