CLONING AND COMPARISON OF A-ALPHA MATING-TYPE ALLELES OF THE BASIDIOMYCETE SCHIZOPHYLLUM-COMMUNE
CLONING AND COMPARISON OF A-ALPHA MATING-TYPE ALLELES OF THE BASIDIOMYCETE SCHIZOPHYLLUM-COMMUNE
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DOI：
10.1007/bf00330567
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发表时间：
1989-07-01
期刊：
影响因子：
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通讯作者：
ULLRICH, RC
中科院分区：
文献类型：
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作者：
GIASSON, L;SPECHT, CA;ULLRICH, RC
A α交配型等位基因（A α 4）通过从紧密连锁的PAB1基因中走染色体而分离。将从步移中分离的含有A α 4等位基因的粘粒克隆用作探针，以从另一粘粒文库中回收A α 1等位基因。通过转化裂褶菌细胞和筛选A调节发育的激活来鉴定编码交配型活性的Cosmetic。在交配试验和后代遗传分析中证实了推定的交配型转化体。分离的特定等位基因的同一性通过显示它们转化交配型的有效性限于具有A α的受体菌株来证明。与克隆序列编码的等位基因不同。转化DNA在反式中是有活性的，表明A α。编码可扩散产物。限制性作图显示A α 1和A α 4在基因组的相同物理区域中编码，但在包含广泛序列差异的亚区域内。此外，Southern分析显示每个单倍体基因组只有一个拷贝的A α 1或A α 4，并且它们彼此之间或与任何其它A α 4不交叉杂交。等位基因。将A α 1和A α 4分别亚克隆为2.8和1.2kb片段，在转化中保留了原始cosplay所证明的所有交配型活性。
An A.alpha. mating-type allele (A.alpha.4) was isolated by walking the chromosome from the closely linked PAB1 gene. A cosmid clone containing the A.alpha.4 allele isolated from the walk was used as a probe to recover the A.alpha.1 allele from another cosmid library. Cosmids encoding mating-type activity were identified by transforming Schizophyllum cells and screening for activation of A-regulated development. Putative mating-type transformants were confirmed in mating tests and genetic analyses of progeny. The identity of the specific alleles isolated was demonstrated by showing that their effectiveness in transforming for mating type is limited to recipient strains possessing an A.alpha. allele different from the one encoded by the cloned sequences. Transforming DNA is active in trans, suggesting the A.alpha. encodes a diffusible product. Restriction mapping shows that A.alpha.1 and A.alpha.4 are coded in the same physical region of the genome, but within a subregion that contains extensive sequence divergence. In addition, Southern analyses show that there is only one copy of A.alpha.1 or A.alpha.4 per haploid genome, and that they do not cross-hybridize to one another or to any of the other A.alpha. alleles. A.alpha.1 and A.alpha.4 were subcloned as 2.8 and 1.2 kb fragments, respectively, retaining in transformation all the mating-type activity demonstrated of the original cosmids.