Biotechnology Fan - Locatinggenesonchromosomes 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 •C o m p l e t e set of mouse chromosome and few human chromosomes Somatic cell hybrid •Fu
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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 06/08/2011 for the course PCB 3063 taught by Professor Marta during the Spring '08 term at University of Florida.

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Biotechnology Fan - Locatinggenesonchromosomes 1....

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