Orr W C, Timberlake W E
Proc Natl Acad Sci U S A. 1982 Oct;79(19):5976-80. doi: 10.1073/pnas.79.19.5976.
We have investigated the chromosomal organization of genes that are expressed specifically in the asexual spores (conidia) of the Ascomycete fungus Aspergillus nidulans, using two experimental approaches. In the first, 30 different recombinant clones, containing long nuclear DNA inserts and at least one spore-specific gene, were selected randomly. The total number of spore-specific genes present in each clone was then determined by RNA blot analysis. In the second approach, several chromosomal recombinant DNA libraries, having average insert lengths ranging from 1 to 15 kilobases, were constructed. The fraction of clones in each library having one or more spore-specific poly(A)+RNA-coding regions was then determined by colony or plaque filter hybridization with radiolabeled, spore-specific, complementary DNA. The results from these experiments were compared to statistical predictions based on the assumption that the spore-specific genes are randomly distributed in the Aspergillus genome. In both cases, the experimental values deviated significantly from the predicted values, demonstrating that the spore-specific genes are nonrandomly arranged in the genome. Rather, they appear frequently to occur in tightly linked clusters.
我们采用两种实验方法,研究了在子囊菌烟曲霉无性孢子(分生孢子)中特异性表达的基因的染色体组织。第一种方法是,随机挑选30个不同的重组克隆,这些克隆含有长核DNA插入片段且至少有一个孢子特异性基因。然后通过RNA印迹分析确定每个克隆中存在的孢子特异性基因的总数。第二种方法是构建几个染色体重组DNA文库,其平均插入片段长度在1至15千碱基之间。然后通过与放射性标记的孢子特异性互补DNA进行菌落或噬菌斑滤膜杂交,确定每个文库中具有一个或多个孢子特异性聚腺苷酸加尾RNA编码区的克隆比例。将这些实验结果与基于孢子特异性基因在曲霉基因组中随机分布这一假设的统计预测结果进行比较。在这两种情况下,实验值均与预测值有显著偏差,表明孢子特异性基因在基因组中的排列是非随机的。相反,它们似乎经常紧密连锁成簇出现。