The establishment and application of positional cloning method for unknown gene in fish species of tetraploid origin.

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As a first step towards the positional cloning of the gene responsible for autosomal dominant oculocutaneous albinism (OCA) in rainbow trout, we performed linkage analysis of SSLP147 markers associated with the dominant albino locus by using 1004 samples from analyzed family. And then, we found that the genetic distance between SSLP147 marker and the dominant albino locus was only 0.2 cM. As a second step, we started chromosome walking method using rainbow trout BAC libraries. As a results, we detected some BAC clones that have no recombinant between phenotypes and genotypes within 1004 samples. We determined whole sequences of the BAC clones and searched the candidate gene in the sequence. Unfortunately, we could not find the candidate gene of OCA. By candidate gene approach method, we isolated rainbow trout c-kit like gene that related with the pigment pattern mutant in zebrafish. We found some base pair substitutions in transcriptional regulatory region, but not in coding region.