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Biotechnology Fan - Locating genes on chromosomes 1...

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Locating genes on chromosomes 1. Fluorescent in situ hybridization (FISH) 2. Somatic cell hybridization
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1. Fluorescent In Situ Hybridization (FISH) Using fluorescent probes that bind to only those parts of the chromosome with which they show a high degree of sequence similarity.
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Method Fix chromosomes to a slide Denature DNA by immersion in a strong base Hybridize probe and chromosomes Visualize probe with fluorescent microscope A human telomere probe to the Pacific oyster ( Crassostrea gigas)
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18S-28S rRNA genes (yellow) Kazuhiko Yamada, Chizuko Nishida-Umehara, Yoichi Matsuda, Molecular and cytogenetic characterization of site-specific repetitive DNA sequences in the Chinese soft-shelled turtle ( Pelodiscus sinensis , Trionychidae), Chromosome Research, Volume 13, Issue 1, Jan 2005, Pages 33 - 46
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2. Somatic Cell hybridization Somatic – body cell (not a gamete, sex cell) Hybridization – two non-homologous cells Human cell Mouse cell Grow together in the presence of inactive Sendai virus Promotes fusion of cells and nuclei
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Single nucleus (unstable) Random loss of human chromosomes Complete set of mouse chromosome and few human chromosomes Somatic cell hybrid Fusion of cells
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Grow different cell lines each with different human chromosomes lost Screen cells to determine which human chromosomes are left
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