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Title: Transgenes of the human hypervariable minisatellite MS32.
Authors: Allen, Maxine Johanna.
Award date: 1994
Presented at: University of Leicester
Abstract: A high rate of mutation at some minisatellite loci can result in exceptional allelic variability. Often, in addition to length polymorphism, sequence variation exists between the minisatellite repeat units in an allele. The interspersion patterns of minisatellite variant repeat units can be determined using MVR-PCR and in this work the allelic hypervariability observed in earlier studies of Ae MS32 locus in Caucasians was shown to extend to Asian alleles. Previous analysis of changes in mutant MS32 MVR-PCR patterns, revealed a polarity of mutation at one end of the repeat array, implicating the possible influence of cis-acting factors on mutation at the MS32 locus. Transgenic mice were created carrying an MS32 transgene to assess for effects of flanking DNA and transgene structure on MS32 mutation processes. Eight separate insertion events were characterised, five single-copy and three multi-copy, and a 'preference' for integration into mouse gamma satellite DNA was observed. Southern blot pedigree analysis and SP-PCR of sperm DNA were performed to assay for mutations at the transgenic loci. Mutation rates at the single-copy loci were 10 to 100-fold lower than at the endogenous human locus. MVR-PCR analysis of SP-PCR recovered transgene 110D mutation events indicated regions of repeat unit duplication, deletion and switching, and a gene conversion event was recovered at the transgene HOC locus. In contrast, mutation rates at the multicopy loci were high (5 - 9.8%) and unusual types of minisatellite mutation including early embryonic events, the frequent loss of a single restriction site and the common deletion of the entire locus were observed. Environmental and genotypic effects on minisatellite mutation were also studied using SP-PCR on DNA from transgene m 110D positive mice irradiated with 60Co, and assaying for minisatellite instability in human cell line DNA with a aberrant mismatch repair phenotype.
Type: Thesis
Level: Doctoral
Qualification: Ph.D.
Rights: Copyright © the author. All rights reserved.
Appears in Collections:Theses, Dept. of Genetics
Leicester Theses

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