Problem
You are using Drosophila as a model system to investigate eye development. You do positional cloning to isolate a mutant gene called eyeless (ey!), which causes flies to develop with reduced sized eyes when homozygous. Using several DNA markers from the candidate interval as probes for colony hybridization to a cDNA library, you obtain a 10 Kb cDNA insert cloned into the unique EcoRI site of a 3.5 Kb plasmid vector.
1. Briefly describe how would you use the Drosophila eyl cDNA clone to identify a clone from a mouse genomic DNA library that is homologous to the Drosophila eyl gene?
2. You isolate a clone of mouse genomic DNA that appears to be homologous to the Drosophila eyl gene After DNA sequencing of a small piece of the mouse genomic clone, you use the DNA sequence to search the mouse genome database. You find that there is a SNP locus (n1234) that does NOT cause an RFLP in this genomic clone. You have identified several breeding lines of mice, each having been bred for several generations, in which abnormal eye development is transmitted as an autosomal recessive condition. How could you test whether mutation of the ey/ gene might cause the abnormal eye development in these lines of mice? Please be sure to explain how you would detect the SNP.
3. Describe two additional lines of evidence that would support the hypothesis that mutation of eyl causes the abnormal eye development phenotype in mice.