Isolation of a Genetic Marker Linked to theBhGene by Genetically Directed Representational Difference Analysis of Closed Colony Japanese Quails

Abstract

Abstract Disappearance of the coloration pattern of the plumage and whole body hemorrhage is caused by an autosomal dominant black at hatch (Bh) mutation in the Japanese quail. In this study, we adopted genetically directed representational difference analysis (GDRDA) to obtain a genetic marker linked to Bh from our closed colony stock. A DNA pool of 10 wild-type (+/+) quails was subtracted from another DNA pool of 10 quails heterozygous for the Bh mutation (Bh/+). After analysis of 106 clones isolated by three series of GDRDA using three different restriction enzymes, one clone was shown to detect a restriction fragment length polymorphism (RFLP) between +/+ and Bh/+ quails. A genomic region flanking the polymorphic restriction site was cloned by inverse-PCR, and a PCR-RFLP maker linked to the Bh mutation was developed. The marker showed no recombination with the Bh mutation in 101 quails analyzed. This is the first report in which a genetic marker linked to a specific gene was successfully isolated by G...