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Deletion Mapping: Closing the gap Chromosome aberrations provide shortcuts to mapping Deletions are particularly useful Principle: a deletion heterozygote with a single copy of the mutant allele should express the phenotype if the gene maps within the deletion complex Deletion mapping the Y chromosome Identify males with cytologically aberrant Y chromosomes Test each male for presence or absence of sequence known to map to the Y chromosome Order the results into a conservative map (the deletion map) Inserting SRY into the deletion map of the Y chromosome Test individuals with “odd” sex: – XY females (lack SRY and define portion of Y not required for sex determination) – XX males (have acquired SRY and define portion of Y essential for sex determination) Screen libraries to conduct “chromosome walking” from known sequences Test retrieved candidate sequences for biological criteria Two important questions in gene cloning How do you get there? – SRY: deletion mapping and chromosome walking How do you know you are there? – SRY: evolutionary conservation in males (“Noah’s ark” blots), interesting sequence, function proven by transgenesis (mouse Sry gene in XX will cause sex reversal to male) DMD: Another monument to deletion cloning Duchenne’s muscular dystrophy known to be X-linked Regional mapping accomplished by two chromosome aberrations: – deletion of DMD and several other genes – Translocation interrupting the DMD gene