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Analysis of Mating-Type Locus Organization and Synteny in Mushroom Fungi: Beyond Model Species

In this chapter genes that are in or near the MAT locus but do not function in determining mating-type specificity are referred to as non-mating-type MAT-linked genes. When trying to study the MAT genes of mushroom fungi other than Coprinopsis cinerea and Schizophyllum commune, there are three major obstacles. First, the loci are likely to be complex and may comprise several genes. Second, the loci display high sequence variation among MAT alleles and cannot be cloned by heterologous hybridization using probes from C. cinerea or S. commune. Finally, the absence of a transformation system in which to test the cloned fragments prevents definitive proof that an isolated DNA region carries MAT. The chapter focuses on how MAT genes may be cloned in nonmodel mushroom species. Although most of this chapter has focused on how conserved gene order can be used to clone MAT from nonmodel species, a more direct option is through PCR amplification of the MAT homologues themselves. The authors have targeted the few conserved amino acids of the homeodomain region (HD1) by degenerate PCR and isolated the entire HD1 gene from this species by genome walking using cassette-mediated PCR.