Flow cytometry isolation and improved visualization of sorted mouse chromosomes

Baron, Bruno; Metezeau, Philippe; Kelly, Françoise; Bernheim, Alain; Berger, Roland; Guénet, Jean-Louis; Goldberg, Michel

doi:10.1016/0014-4827(84)90247-7

Cited by 28 publications

(31 citation statements)

References 10 publications

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“…Following stringent washes and autoradiography, one positive BAC (272J2) was identi®ed, and characterized by STS content and restriction cleavage (to be detailed elsewhere). The BAC was biotin-labelled and used as a probe for FISH analysis as described (The European Consortium on MEN1, 1996) on metaphase chromosomes from mouse SV22CD cells (Baron et al, 1984). Fluorescence images were captured using a high-performance cooled chargecoupled device (CCD) camera C4880 (Hamamatsu), interfaced to a PC 486DX33 with a Matrox 640 card.…”

Section: Chromosome Mappingmentioning

confidence: 99%

Characterization of the mouse Men1 gene and its expression during development

et al. 1998

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The gene responsible for multiple endocrine neoplasia type 1 (MEN1), a heritable predisposition to endocrine tumours in man, has recently been identi®ed. Here we have characterized the murine homologue with regard to cDNA sequence, genomic structure, expression pattern and chromosomal localisation. The murine Men1 gene spans approximately 6.7 kb of genomic DNA and is comprised of 10 exons with similar genomic structure to the human locus. It was mapped to the pericentromeric region of mouse chromosome 19, which is conserved with the human 11q13 band where MEN1 is located. The predicted protein is 611 amino acids in length and overall is 97% homologous to the human orthologue. The 45 reported MEN1 mutations which alter or delete a single amino acid in human all occur at conserved residues, thereby supporting their functional signi®cance. Two transcripts of approximately 3.2 and 2.8 kb were detected in both embryonal and adult murine tissues, resulting from alternative splicing of intron 1. By RNA in situ hybridization and Northern analysis the spatiotemporal expression pattern of Men1 was determined during mouse development. Men1 gene activity was detected already at gestational day 7. At embryonic day 14 expression was generally high throughout the embryo, while at day 17 the thymus, skeletal muscle, and CNS showed the strongest signal. In selected tissues from postnatal mouse Men1 was detected in all tissues analysed and was expressed at high levels in cerebral cortex, hippocampus, testis, and thymus. In brain the menin protein was detected mainly in nerve cell nuclei, whereas in testis it appeared perinuclear in spermatogonia. These results show that Men1 expression is not con®ned to organs a ected in MEN1, suggesting that Men1 has a signi®cant function in many di erent cell types including the CNS and testis.

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Section: Chromosome Mappingmentioning

confidence: 99%

Characterization of the mouse Men1 gene and its expression during development

et al. 1998

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“…In the karyotype of the SV 22CD cell line, all chromosomes are fused in pairs, except for chromosome 19 and the slightly larger X chromosome (8). This chromosomal abnormal- Cig30 and Pitx3 Overlapity was accidentally of great benefit in the chromosomal determination, because the twin spot signals were observed specifically on a single unfused chromosome pair, which was readily identified as chromosome 19 ( Fig.…”

Section: Isolation Of Mouse Cig30mentioning

confidence: 96%

“…100 ng of labeled DNA was coprecipitated in the presence of 20 g of Cot1 DNA (Life Technologies, Inc.), denatured at 70°C for 10 min, and pre-annealed at 37°C for 30 min. An overnight hybridization was then carried out on metaphase chromosomes from the mouse SV 22CD cell line (8). The signal was detected using fluorescein isothiocyanate-conjugated avidin (Vector Laboratories).…”

Section: Methodsmentioning

confidence: 99%

Cig30 and Pitx3 Genes Are Arranged in a Partially Overlapping Tail-to-Tail Array Resulting in Complementary Transcripts

Tvrdík

Asadi

Kozak

et al. 1999

Journal of Biological Chemistry

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“…Thirteen populations have been resolved and sorting for confirmation of peak Identity is underway (Cram, 1992). Alternative approaches for resolving the mouse karyotype have been demonstrated using mouse cell lines with multiple Robertsonian translocations (Baron, 1984, Baron, 1986. Several groups are involved in porcine chromosome sorting for the Pig Gene Mapping Project (Haley, 1990).…”

Section: 'Fluorescence Intensitymentioning

confidence: 99%