Specht C A, Stankis M M, Novotny C P, Ullrich R C
Department of Botany, University of Vermont, Burlington 05405.
Genetics. 1994 Jul;137(3):709-14. doi: 10.1093/genetics/137.3.709.
Classical genetic studies identified nine mating-type specificities at the A alpha locus of the Basidiomycete fungus Schizophyllum commune. We have used Southern blot hybridizations to generate EcoRI restriction maps of the A alpha locus for 18 strains, including all nine specificities. A alpha 1, A alpha 3 and A alpha 4 DNA was subcloned from three cosmids and used as probes. A unique region of DNA was found for each of the three cloned specificities. Hybridization was detected in this region only if the probe(s) and the blotted genomic DNAs were from strains with the same A alpha specificity. DNAs from strains with the same A alpha specificity hybridize regardless of geographic origin, but DNAs from strains with different A alpha specificities do not cross-hybridize. The results demonstrate two size classes of unique A alpha DNA. This unique DNA is about 4.5 kb in A alpha 1 strains and about 7.0-8.5 kb in other strains. Transcription regulators Z and Y, which were deduced previously from the DNA sequence of the A alpha 1, A alpha 3 and A alpha 4 loci, are probably encoded by all non-A alpha 1 loci. The smaller A alpha 1 loci appear to encode only Y and lack sequence for Z. No evidence was found for a locus that encodes only Z. The lack of hybridization detected between A alpha loci with different specificities suggests that the evolution of A alpha has resulted from extensive sequence divergence.
经典遗传学研究在担子菌裂褶菌的Aα位点鉴定出了9种交配型特异性。我们利用Southern印迹杂交技术,为18个菌株(包括所有9种特异性类型)构建了Aα位点的EcoRI限制酶切图谱。从三个黏粒中分别亚克隆出Aα1、Aα3和Aα4 DNA,并用作探针。对于这三种克隆的特异性类型,均发现了一个独特的DNA区域。只有当探针和印迹的基因组DNA来自具有相同Aα特异性的菌株时,才会在该区域检测到杂交信号。来自具有相同Aα特异性的菌株的DNA,无论其地理来源如何,都会发生杂交,但来自具有不同Aα特异性的菌株的DNA不会发生交叉杂交。结果表明存在两种大小类别的独特Aα DNA。这种独特的DNA在Aα1菌株中约为4.5 kb,在其他菌株中约为7.0 - 8.5 kb。先前从Aα1、Aα3和Aα4位点的DNA序列推导出来的转录调节因子Z和Y,可能由所有非Aα1位点编码。较小的Aα1位点似乎只编码Y,而缺少Z的序列。未发现仅编码Z的位点的证据。不同特异性的Aα位点之间未检测到杂交,这表明Aα的进化是由广泛的序列分歧导致的。