Rapid fluorescencein situ hybridization on interphasic nuclei to discriminate between homozygous and heterozygous transgenic mice |
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Authors: | Daniel Paris Kiyoko Toyama André Mégarbane Myriam Casanova Pierre-Marie Sinet Jacqueline London |
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Affiliation: | (1) CNRS URA 1335, Faculté de Médecine Necker-Enfants Malades, 156 rue de Vaugirard, 75730 Paris Cedex 15, France;(2) Service d'Histologie, Embryologie et de Cytogénétique, Hopital Necker-Enfants Malades, 149 rue de Sèvres, 75743 Paris Cedex 15, France |
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Abstract: | Homozygous and heterozygous transgenic mice of the Tg152 line overexpressing the human copper/zinc superoxide dismutase (hSOD-1) were rapidly differentiated by fluorescencein situ hybridization (FISH) using intérphase lymphocyte nuclei. We have devised a simple and fast method for preparing interphase nuclei with very small quantities of whole mouse blood, avoiding several steps of the classical FISH technique. Lymphocyte separation and cell culture were not required. This technique provides an excellent tool for the unambiguous detection of homozygous and heterozygous transgenic mice in a litter. It can be used to check young animals since 2 l of whole blood is sufficient. We also show that in this transgenic line numerous copies of the hSOD-1 transgene are. integrated at a single autosomal locus, in tandem head-to-tail organization |
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Keywords: | FISH on mouse interphasic nuclei transgenic inbred FVB/N hSOD-1 mice rapid hSOD-1 assay |
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