Biotechnology Fan - Locating genes on chromosomes 1....

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Unformatted text preview: Locating genes on chromosomes 1. Fluorescent in situ hybridization (FISH) 2. Somatic cell hybridization 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. 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) 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 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 • Single nucleus (unstable) • Random loss of human chromosomes • Complete set of mouse chromosome and few human chromosomes Somatic cell hybrid • Fusion of cells • Grow different cell lines each with different human chromosomes lost • Screen cells to determine which human chromosomes are left • Probe for gene of interest in each cell line...
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This note was uploaded on 12/02/2011 for the course PCB 3063 taught by Professor Marta during the Fall '08 term at University of Florida.

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Biotechnology Fan - Locating genes on chromosomes 1....

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