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Mammalian Genome

, Volume 13, Issue 12, pp 680–685 | Cite as

Rapid localization of transgenes in mouse chromosomes with a combined Spectral Karyotyping/FISH technique

  • Sei-ichi Matsui
  • Sheila Sait
  • Craig A. Jones
  • Norma Nowak
  • Kenneth W. Gross

We explored the feasibility of combined Spectral Karyotyping (SKY) and Fluorescence In Situ Hybridization (FISH) as means to rapidly map a chromosomally integrated renin/green fluorescent protein (GFP) fusion gene construct (Ren-GFP) in the transgenic mouse, Tg(Ren-GFP)1Kwg. A sequential hybridization with SKY probes followed by FISH gave consistently satisfactory results, demonstrating that multiple copies of the Ren-GFP transgene in this transgenic mouse line are integrated into a single chromosomal site of Chromosome (Chr) 4, most probably in the juxta-centromeric euchromatic region consisting of the A2-A3 domain. Chr 4 as a sole carrier of the transgene also was confirmed by co-hybridization to p1 BAC clone DNA containing telomeric sequences specific for mouse Chr 4 and the Ren-GFP construct in pGEM4Z vector. The hemizygosity of the Ren-GFP transgene is maintained not only in bone marrow cells, but also in lung cells proliferating in vitro, indicating that stable integration of the Ren-GFP transgene into chromosomal DNA was established at a very early embryonic stage. We conclude that the SKY/FISH technique is a reliable and facile method for establishing the integration site of a transgene. As such, this protocol has obvious advantages over traditional backcross methods in terms of time, cost and labor for determining the chromosomal location of transgenes.

Keywords

Telomeric Sequence Transgenic Mouse Line Euchromatic Region Chromosomal Site Early Embryonic Stage 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag New York Inc. 2002

Authors and Affiliations

  • Sei-ichi Matsui
    • 2
  • Sheila Sait
    • 2
  • Craig A. Jones
    • 3
  • Norma Nowak
    • 1
  • Kenneth W. Gross
    • 3
  1. 1.SKY and DNA Microarray Laboratories, Dept. of Cancer Genetics, Roswell Park Cancer Institute, Buffalo, New York 14263, USAUS
  2. 2.Clinical Cytogenetics, Dept. of Pathology and Laboratory Medicine, Roswell Park Cancer Institute, Buffalo, New York 14263, USAUS
  3. 3.Dept. of Molecular and Cellular Biology, Roswell Park Cancer Institute, Buffalo, New York 14263, USAUS

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