BIOCHEMICAL AND SOMATIC CELL GENETIC STUDIES OF HUMAN HEME BIOSYNTHETIC ENZYMES.
Item
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Title
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BIOCHEMICAL AND SOMATIC CELL GENETIC STUDIES OF HUMAN HEME BIOSYNTHETIC ENZYMES.
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Identifier
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AAI8312348
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identifier
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8312348
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Creator
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GIAMPIETRO, PHILIP FRANCIS.
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Contributor
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R.J. Desnick
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Date
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1983
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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, Genetics
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Abstract
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The regional gene assignment for human porphobilinogen deaminase (PBG-deaminase; E.C.4.3.18) on chromosome 11 has been determined using somatic cell hybridization, immunologic, electrophoretic, and cytogenetic techniques. Dimethyl sulfoxide (DMSO) -induced erythroid differentiation of hybrid clones derived from the fusion of tetraploid Friend murine erythroleukemia (2S MEL) cells deficient in thymidine kinase and human Lesch-Nyhan fibroblasts (HLN) deficient in hypoxanthine phosphoribosyltransferase were examined for the expression of human PBG-deaminase, esterase A(,4) and lactate dehydrogenase A. Examination of five primary and 10 secondary, tertiary or quaternary clones permitted the regional assignment of human PBG-deaminase to the long arm of chromosome 11. The analysis of two 2S MEL (HPRT('-))-human fibroblast (HX/11) hybrids, each containing the human X chromosome-autosome translocation (der 11), t(X;11)(q25-26; q23) as the only human chromosome permitted finer localization of the gene for human PGB-deaminase to the region 11q23 (--->) 11qter.;Twelve RAG cell-human fibroblast cell hybrids were examined by electrophoretic and immunologic methods for the presence of human (delta)-aminolevulinic acid dehydratase (ALA-dehydratase). Using these methods it was not possible to detect human ALA-dehydratase, although the mouse enzyme was electrophoretically and immunologically detectable. Possible reasons for the lack of expression of human ALA-dehydratase are discussed.;Further studies concerning ALA-dehydratase included the development of a sensitive fluorometric assay for determining enzymatic activity in human fibroblasts and amniocytes. Naturally occurring ALA-dehydratase polymorphisms were further characterized with respect to electrophoretic mobility, binding to DEAE-cellulose, K(,m) and thermostability properties. ALA-dehydratase protein levels were determined in erythrocyte lysates from normal and ALA-dehydratase deficient individuals by rocket immunoelectrophoresis. Erythrocyte lysates obtained from heterozygotes for ALA-dehydratase deficiency were found to contain normal levels of ALA-dehydratase immunoreactive protein.
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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
