MUTAGENESIS AND ALU SEQUENCE-MEDIATED DNA DISPERSION IN TRANSGENIC MICE MICROINJECTED WITH HUMAN GLOBIN GENES.
Item
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Title
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MUTAGENESIS AND ALU SEQUENCE-MEDIATED DNA DISPERSION IN TRANSGENIC MICE MICROINJECTED WITH HUMAN GLOBIN GENES.
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Identifier
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AAI8801754
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identifier
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8801754
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Creator
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RUBINSTEIN, WENDY SUE.
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Date
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1987
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Language
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English
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Publisher
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City University of New York.
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Subject
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Biology, Molecular
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Abstract
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Human globin genes were microinjected to produce transgenic mice, and two lines microinjected with the phage clone {dollar}\lambda{dollar}H{dollar}\beta{dollar}G1 were further evaluated. Line {dollar}\beta{dollar}80, which carries 50-100 copies of the foreign construct in tandem array, displayed pleiotropic mutations. Heterozygous mice developed lymphoma and leukemia at 13-16 months of age, and some mice displayed infertility and neurologic defects. Homozygosity of the foreign insert caused an embryonic lethal mutation which was about 80% penetrant. Rare surviving homozygous mice, whose genotypes were evaluated by southern blotting intensities, showed abnormal forelimb development and overall growth retardation, and usually died within a few days. In addition, a sex ratio distortion was observed in the {dollar}\beta{dollar}80 line. The locus of insertion was isolated in cosmid cloning vectors, and subclones were used to search for restriction fragment-length polymorphisms and for chromosomal mapping. Studies in progress will determine if this line carries an analogous biochemical defect to that seen in the human chromosome breakage syndrome, Franconi's anemia.;In line {dollar}\beta{dollar}19, southern blot analysis on the spleen of the founder mouse revealed an unusual hybridization pattern which suggested extensive dispersion of human DNA throughout the mouse genome. This pattern was reproducible using several restriction enzymes including a non-cutting enzyme. The hybridization pattern was not observed in other tissues, and sequences were not detected in progeny using the bacteriophage probe. However, hybridization of spleen DNA of offspring against a human Alu probe revealed genetic transmission of human Alu sequences. The results suggest dispersion of microinjected human Alu sequences throughout the mouse genome by a mechanism which is currently under investigation.
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Type
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dissertation
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Source
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PQT Legacy CUNY.xlsx
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degree
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Ph.D.
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Program
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Biomedical Sciences
