You obtain a mouse mutant in which the skin is severely blistered. The mutation

Question

You obtain a mouse mutant in which the skin is severely blistered. The mutation maps near the gene encoding the alpha6 integrin protein. a. Why would a mutation in integrin cause blistering of the skin? What other proteins would you expect to produce this phenotype when mutated and why? b. You make a cell line from the skin epithelial cells of this mutant mouse. What are two methods you could use with these cells to test whether the mutant phenotype could be the consequence of mutation of this integrin gene? Explain the predicted results for each approach.

Explanation / Answer

A ) If the epithelial cells are not attached to the basal lamina the space between the two fills with fluid to provide blisters. Mutation of any of the additives of the basal lamina accountable for attachment to the epithelial cells could produce the same phenotype. The most likely candidate could be laminin, but additionally mutation of collagen IV or the proteoglycans might disrupt basal lamina shape sufficient that laminin couldn't bind integrin at the epithelial mobile nicely.


B) 1. You ought to isolate genomic DNA, PCR enlarge the alpha6 integrin gene and collection it. Finding a mutation within the gene might suggest that this gene changed into answerable for the mutant phenotype.

2. You should bind the cells with an antibody in opposition to alpha6 integrin and see if it changed into at the cellular surface within the mutant cells. If it have been no longer that might provide an explanation for the phenotype. You may also use your antibody to study the ranges or mobility types of the protein on westerns to look in the event that they have been altered within the mutant cells.

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