Identification of the albino mutation of mouse tyrosinase by analysis of an in vitro revertant

Abstract

From within an albino melanocyte line grown in vitro we identified and cloned cells that apparently had reverted to wild type. We sequenced a part of the tyrosinase gene, encompassing a candidate mutation, from wild-type, albino, and revertant cell DNAs. The revertant cells contain, on one chromosome, a perfect base reversion to the wild-type sequence of this candidate mutation, proving that this is the sole defect in the tyrosinase gene of albino mutant mice. The revertant cells readily regain the albino phenotype after freezing and thawing. Taking advantage of a Dde I restriction site created by the albino mutation, we demonstrated that the regained phenotype is due to allele loss involving the wild-type chromosome. The Dde I site also allowed us to show that all inbred albino mice carry the same mutation and so must be derived from the same progenitor.