Construction and characterization of a DNA library from mouse chromosomes 19 purified by flow cytometry.
In spite of the constant development concerning physical mapping of eukaryotic genomes, the mouse chromosome 19 remains poorly characterized. In order to improve the possibilities for studying this chromosome, we have constructed a chromosome-specific EcoRI DNA fragment library from mouse chromosomes 19 sorted by flow cytometry. The resulting library contains about 3 X 10(4) recombinant clones. The identified inserts range in size from about 0.2-10 kb, with a 4 kb average size and with no observable redundancy. The purity of the library has been analyzed by flow-blot. For that purpose, chromosomes from 2 cell lines, 1 with a normal karyotype and 1 with translocated chromosome 19, were sorted on nylon filters and hybridized with 9 clones of the library. Results show that 5 clones out of the 9 clearly originate from sorted chromosomes 19 and 3 and are likely to be derived from its DNA, thus indicating that the library of chromosome 19 is of high purity.
Baron
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B
Service commun INSERM-Pasteur Trieur de Cellules, Unité de Biochimie Cellulaire, Institut Pasteur, Paris, France.
Métézeau
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P
Kiefer-Gachelin
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H
Goldberg
M E
ME
D000818:Animals; D010582:Bacteriophage lambda; D002458:Cell Fractionation / Q000379:methods; D002875:Chromosomes; D003001:Cloning, Molecular; D005434:Flow Cytometry; D005784:Gene Amplification; D015723:Gene Library; D051379:Mice; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D012091:Repetitive Sequences, Nucleic Acid