Isolation and characterization of a minor domain of human Satellite II DNA.
Item
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Title
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Isolation and characterization of a minor domain of human Satellite II DNA.
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Identifier
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AAI8821103
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identifier
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8821103
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Creator
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Macoska, Jill Anne.
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Contributor
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Adviser: Ann Henderson
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Date
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1988
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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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Chemistry, Biochemistry | Biology, General | Biology, Genetics
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Abstract
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Specific sequence heterogeneities and a novel minor chromosome 7-specific domain were identified within human Satellite II DNA. Satellite II DNA was purified using cesium gradient ultracentrifugation and propagated as recombinant DNAs. Twelve of 106 cloned DNA fragments were randomly chosen for study. Three contained unique or not detectably repetitive sequences. One contained alphoid DNA. Four contained sequences homologous to L1 KpnI LINES element DNA and one of these, 43 DNA, localized specifically to the centromeres of chromosome 4 and the X chromosome. The identification of unique, alphoid and L1 sequences defined nonsatellite-associated sequence heterogeneity within Satellite II DNA. The remaining four cloned DNAs studied contained Satellite II sequences associated with Alu SINES element DNA. DNA base sequencing showed that the 5{dollar}\sp\prime{dollar} terminal 200 basepair (bp) region of one of these, 33 DNA, contained a tandem array of six degenerate 26-nucleotide Satellite II repeat units that conserved an average 56% of the consensus sequence residues. This region was subcloned as XL3 DNA and comprised basepairs 1-750 of 33 DNA. Southern blot analysis showed that a 1400 basepair sequence homologous to XL3 DNA occurred 10-100X in genomic DNA. Moreover, XL3 DNA revealed a 3.4 kilobasepair (kb) HaeIII fragment in human male placental DNA and hybridized in situ to the pericentromeric region of human chromosome 7. This indicated that XL3 DNA was homologous to the non Y-specific (NYS) 3.4 kb HaeIII family of repeated sequences. Together, these experiments showed that XL3 DNA defined a novel minor domain of human Satellite II that occurred in 10-100 copies in genomic DNA, was associated with NYS 3.4 kb HaeIII repeated sequences and localized to the pericentromeric region of human chromosome 7.
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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.
