Zimmermann C R, Orr W C, Leclerc R F, Barnard E C, Timberlake W E
Cell. 1980 Oct;21(3):709-15. doi: 10.1016/0092-8674(80)90434-1.
Over 350 clones homologous to poly(A)+ RNAs that are significantly more prevalent in conidiating cultures of Aspergillus nidulans than in somatic cells have been selected from a recombinant DNA library formed between nuclear DNA and lambda Charon 4A. The procedure used for this selection involved in situ hybridization to a cDNA probe which had been selectively depleted of sequences represented in somatic cells by complement hybridization. Five of these clones have been characterized further. All but one encoded poly(A)+ RNAs that were at least ten times more prevalent in conidiating cultures than in somatic cells. One clone hybridized to a single, developmentally regulated RNA. The three others were complementary to several RNAs having different molecular weights, each of which was more prevalent in condiating cultures than in vegetative cells. These results and quantitative aspects of the selection procedure suggest that developmentally controlled poly(A)+ RNA coding regions may not be distributed randomly in the Aspergillus genome.
从构巢曲霉核DNA与λ噬菌体Charon 4A构建的重组DNA文库中,已筛选出350多个与多聚腺苷酸加尾(poly(A)+)RNA同源的克隆,这些克隆在构巢曲霉的分生孢子形成培养物中比在体细胞中更为普遍。用于该筛选的程序包括与一个cDNA探针进行原位杂交,该探针已通过互补杂交选择性地去除了体细胞中存在的序列。其中五个克隆已得到进一步表征。除了一个克隆外,所有其他克隆编码的多聚腺苷酸加尾RNA在分生孢子形成培养物中的丰度至少比在体细胞中高十倍。一个克隆与一种单一的、受发育调控的RNA杂交。另外三个克隆与几种分子量不同的RNA互补,每种RNA在分生孢子形成培养物中的丰度都比在营养细胞中高。这些结果以及筛选程序的定量方面表明,受发育控制的多聚腺苷酸加尾RNA编码区域在构巢曲霉基因组中可能不是随机分布的。