Pate, Lorna Wendy; (1995) Mapping studies of mouse chromosome two. Doctoral thesis (Ph.D), UCL (University College London).

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Abstract

Mouse chromosome two harbours the interesting mutant genes ragged (Ra), wasted (wst), lethal spotting (ls) and ulnaless (Ul). These studies were intended to provide resources for the initiation of positional cloning projects to isolate one or more of the genes involved in these mutations. The first approach was the construction and characterization of two new panels of mouse: Chinese hamster somatic cell hybrids, the mouse parental cell line in each case being derived from spleen cells of mice carrying either the T(2;8)2Wa or the T(2;16)28H reciprocal translocations. These translocation breakpoints flank the region on distal mouse chromosome two in which ragged and wasted are thought to lie, and provide a means of defining the physical limits of the region. Forty four separate cell lines were produced, but no segregation of the translocation reciprocals in either panel was noted, despite subcloning. Forced antibody selection strategies were also investigated. The second aim of the project was to generate new molecular markers for mouse chromosome two by species specific Interspersed Repetitive Sequence Polymerase Chain Reaction (IRS-PCR) and Interspersed Repetitive Sequence to bubble (IRS-bubble) Polymerase Chain Reaction amplification of DNA from hybrid cells containing only mouse chromosome two. The PCR products were screened for the presence of putative CpG islands and microsatellite repeats. Fragments were then cloned and sequenced. PCR primers were designed from the sequence obtained, and variation was identified between Mus musculus and Mus spretus. Twelve new markers were produced. The new markers were then placed on the map of mouse chromosome two by interspecific backcross analysis. New markers showing linkage to ragged, wasted or ulnaless were then mapped in relation to these mutants by the use of interspecific and intersubspecific backcrosses in which the mutant was segregating.

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