Skip to main content
NCBI home page
Genetics logoLink to Genetics
. 1995 Sep;141(1):321–332. doi: 10.1093/genetics/141.1.321

Phenotypic and Molecular Analysis of a Transgenic Insertional Allele of the Mouse Fused Locus

W L Perry-III 1, T J Vasicek 1, J J Lee 1, J M Rossi 1, L Zeng 1, T Zhang 1, S M Tilghman 1, F Costantini 1
PMCID: PMC1206729  PMID: 8536979

Abstract

Spontaneous mutations at the mouse Fused (Fu) locus cause dominant skeletal and neurological defects and recessive lethal embryonic defects including neuroectodermal abnormalities and axial duplications. Here, we describe a new allele at the Fu locus caused by a transgenic insertional mutation, Hε46. Embryos homozygous for the Hε46 insertion die at day 9-10 post coitum and display phenotypic defects similar to those associated with Fu alleles. The Hε46 locus was cloned and shown to contain a 20-kb deletion at the site of transgene insertion with no other detectable rearrangements. Genomic probes from the Hε46 locus were mapped to a genetic locus closely linked to Fu on chromosome 17 and were hybridized to a YAC contig covering the Fu(Ki) critical region. Compound heterozygotes between Hε46 and Fu(Ki) were inviable and displayed abnormalities at the same stage of embryogenesis as do homozygotes for either of the two mutations, demonstrating that these two recessive lethal mutations belong to the same complementation group. A genomic probe from the wild-type Hε46 locus detected a transcript that is disrupted by the transgenic insertion, representing a candidate for the wild-type allele of Fused.

Full Text

The Full Text of this article is available as a PDF (7.7 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Belyaev D. K., Ruvinsky A. O., Borodin P. M. Inheritance of alternative states of the fused gene in mice. J Hered. 1981 Mar-Apr;72(2):107–112. doi: 10.1093/oxfordjournals.jhered.a109436. [DOI] [PubMed] [Google Scholar]
  2. Cattanach B. M. Parental origin effects in mice. J Embryol Exp Morphol. 1986 Oct;97 (Suppl):137–150. [PubMed] [Google Scholar]
  3. Cheng S. S., Costantini F. Morula decompaction (mdn), a preimplantation recessive lethal defect in a transgenic mouse line. Dev Biol. 1993 Mar;156(1):265–277. doi: 10.1006/dbio.1993.1075. [DOI] [PubMed] [Google Scholar]
  4. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  5. Conlon F. L., Lyons K. M., Takaesu N., Barth K. S., Kispert A., Herrmann B., Robertson E. J. A primary requirement for nodal in the formation and maintenance of the primitive streak in the mouse. Development. 1994 Jul;120(7):1919–1928. doi: 10.1242/dev.120.7.1919. [DOI] [PubMed] [Google Scholar]
  6. Copeland N. G., Jenkins N. A. Development and applications of a molecular genetic linkage map of the mouse genome. Trends Genet. 1991 Apr;7(4):113–118. doi: 10.1016/0168-9525(91)90455-y. [DOI] [PubMed] [Google Scholar]
  7. Copeland N. G., Jenkins N. A., Gilbert D. J., Eppig J. T., Maltais L. J., Miller J. C., Dietrich W. F., Weaver A., Lincoln S. E., Steen R. G. A genetic linkage map of the mouse: current applications and future prospects. Science. 1993 Oct 1;262(5130):57–66. doi: 10.1126/science.8211130. [DOI] [PubMed] [Google Scholar]
  8. Costantini F., Radice G., Lee J. L., Chada K. K., Perry W., Son H. J. Insertional mutations in transgenic mice. Prog Nucleic Acid Res Mol Biol. 1989;36:159–169. doi: 10.1016/s0079-6603(08)60169-5. [DOI] [PubMed] [Google Scholar]
  9. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  10. Forejt J., Artzt K., Barlow D. P., Hamvas R. M., Lindahl K. F., Lyon M. F., Klein J., Silver L. M. Mouse chromosome 17. Mamm Genome. 1994;5(Spec No):S238–S258. [PubMed] [Google Scholar]
  11. Green E. D., Olson M. V. Systematic screening of yeast artificial-chromosome libraries by use of the polymerase chain reaction. Proc Natl Acad Sci U S A. 1990 Feb;87(3):1213–1217. doi: 10.1073/pnas.87.3.1213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Greenspan R. J., O'Brien M. C. Genetic analysis of mutations at the fused locus in the mouse. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4413–4417. doi: 10.1073/pnas.83.12.4413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gridley T. Insertional versus targeted mutagenesis in mice. New Biol. 1991 Nov;3(11):1025–1034. [PubMed] [Google Scholar]
  14. Himmelbauer H., Harvey R. P., Copeland N. G., Jenkins N. A., Silver L. M. High-resolution genetic analysis of a deletion on mouse chromosome 17 extending over the fused, tufted, and homeobox Nkx2-5 loci. Mamm Genome. 1994 Dec;5(12):814–816. doi: 10.1007/BF00292022. [DOI] [PubMed] [Google Scholar]
  15. Himmelbauer H., Silver L. M. High-resolution comparative mapping of mouse chromosome 17. Genomics. 1993 Jul;17(1):110–120. doi: 10.1006/geno.1993.1291. [DOI] [PubMed] [Google Scholar]
  16. Hodgkinson C. A., Moore K. J., Nakayama A., Steingrímsson E., Copeland N. G., Jenkins N. A., Arnheiter H. Mutations at the mouse microphthalmia locus are associated with defects in a gene encoding a novel basic-helix-loop-helix-zipper protein. Cell. 1993 Jul 30;74(2):395–404. doi: 10.1016/0092-8674(93)90429-t. [DOI] [PubMed] [Google Scholar]
  17. Jacobs-Cohen R. J., Spiegelman M., Cookingham J. C., Bennett D. Knobbly, a new dominant mutation in the mouse that affects embryonic ectoderm organization. Genet Res. 1984 Feb;43(1):43–50. doi: 10.1017/s0016672300025702. [DOI] [PubMed] [Google Scholar]
  18. King T. R., Dove W. F., Herrmann B., Moser A. R., Shedlovsky A. Mapping to molecular resolution in the T to H-2 region of the mouse genome with a nested set of meiotic recombinants. Proc Natl Acad Sci U S A. 1989 Jan;86(1):222–226. doi: 10.1073/pnas.86.1.222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kusumi K., Smith J. S., Segre J. A., Koos D. S., Lander E. S. Construction of a large-insert yeast artificial chromosome library of the mouse genome. Mamm Genome. 1993;4(7):391–392. doi: 10.1007/BF00360591. [DOI] [PubMed] [Google Scholar]
  20. Lee J. J., Radice G., Perkins C. P., Costantini F. Identification and characterization of a novel, evolutionarily conserved gene disrupted by the murine H beta 58 embryonic lethal transgene insertion. Development. 1992 May;115(1):277–288. doi: 10.1242/dev.115.1.277. [DOI] [PubMed] [Google Scholar]
  21. Lee J. J., Warburton D., Robertson E. J. Cytogenetic methods for the mouse: preparation of chromosomes, karyotyping, and in situ hybridization. Anal Biochem. 1990 Aug 15;189(1):1–17. doi: 10.1016/0003-2697(90)90036-9. [DOI] [PubMed] [Google Scholar]
  22. Lyon M. F., Bechtol K. B. Derivation of mutant t-haplotypes of the mouse by presumed duplication or deletion. Genet Res. 1977 Aug;30(1):63–76. doi: 10.1017/s0016672300017468. [DOI] [PubMed] [Google Scholar]
  23. Radice G., Lee J. J., Costantini F. H beta 58, an insertional mutation affecting early postimplantation development of the mouse embryo. Development. 1991 Mar;111(3):801–811. doi: 10.1242/dev.111.3.801. [DOI] [PubMed] [Google Scholar]
  24. Riley J., Butler R., Ogilvie D., Finniear R., Jenner D., Powell S., Anand R., Smith J. C., Markham A. F. A novel, rapid method for the isolation of terminal sequences from yeast artificial chromosome (YAC) clones. Nucleic Acids Res. 1990 May 25;18(10):2887–2890. doi: 10.1093/nar/18.10.2887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Robertson E., Bradley A., Kuehn M., Evans M. Germ-line transmission of genes introduced into cultured pluripotential cells by retroviral vector. Nature. 1986 Oct 2;323(6087):445–448. doi: 10.1038/323445a0. [DOI] [PubMed] [Google Scholar]
  26. Rossi J. M., Burke D. T., Leung J. C., Koos D. S., Chen H., Tilghman S. M. Genomic analysis using a yeast artificial chromosome library with mouse DNA inserts. Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2456–2460. doi: 10.1073/pnas.89.6.2456. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rossi J. M., Chen H., Tilghman S. M. Genetic map of the fused locus on mouse chromosome 17. Genomics. 1994 Sep 1;23(1):178–184. doi: 10.1006/geno.1994.1475. [DOI] [PubMed] [Google Scholar]
  28. Ruvinsky A. O., Agulnik A. I. Gametic imprinting and the manifestation of the fused gene in the house mouse. Dev Genet. 1990;11(4):263–269. doi: 10.1002/dvg.1020110404. [DOI] [PubMed] [Google Scholar]
  29. Ruvinsky A., Agulnik A., Agulnik S., Rogachova M. Functional analysis of mutations of murine chromosome 17 with the use of tertiary trisomy. Genetics. 1991 Apr;127(4):781–788. doi: 10.1093/genetics/127.4.781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Woychik R. P., Maas R. L., Zeller R., Vogt T. F., Leder P. 'Formins': proteins deduced from the alternative transcripts of the limb deformity gene. Nature. 1990 Aug 30;346(6287):850–853. doi: 10.1038/346850a0. [DOI] [PubMed] [Google Scholar]
  32. Woychik R. P., Maas R. L., Zeller R., Vogt T. F., Leder P. 'Formins': proteins deduced from the alternative transcripts of the limb deformity gene. Nature. 1990 Aug 30;346(6287):850–853. doi: 10.1038/346850a0. [DOI] [PubMed] [Google Scholar]
  33. Xiang X., Benson K. F., Chada K. Mini-mouse: disruption of the pygmy locus in a transgenic insertional mutant. Science. 1990 Feb 23;247(4945):967–969. doi: 10.1126/science.2305264. [DOI] [PubMed] [Google Scholar]
  34. Zhou X., Sasaki H., Lowe L., Hogan B. L., Kuehn M. R. Nodal is a novel TGF-beta-like gene expressed in the mouse node during gastrulation. Nature. 1993 Feb 11;361(6412):543–547. doi: 10.1038/361543a0. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES