Clancy M J, Buten-Magee B, Straight D J, Kennedy A L, Partridge R M, Magee P T
Proc Natl Acad Sci U S A. 1983 May;80(10):3000-4. doi: 10.1073/pnas.80.10.3000.
A library of Saccharomyces cerevisiae DNA in the vector lambda Charon 28 was probed for sequences complementary to cDNA made from poly(A)+ RNA isolated from the well-sporulating yeast strain AP1 a/alpha. The RNA was isolated from cells that had been incubated 7, 9, 11, and 13 hr in sporulation medium. DNA complementary to poly(A)+ RNA from alpha/alpha(nonsporulating) AP1 was used as a control, and 46 bacteriophage that gave a stronger response with a/alpha cDNA than with alpha/alpha cDNA were obtained in a screening of three yeast genomes worth of DNA. Two of the bacteriophage appeared to contain a/alpha-specific genes, in that they hybridized to cDNA from vegetative a/alpha RNA. The rest appeared to correspond to a/alpha genes expressed preferentially during sporulation. Restriction endonuclease analysis of four of the cloned sequences revealed a single major region of transcription in each; these regions ranged in size from 2.5 to 4.0 kilobases. RNA blot analysis showed that, in three of the four cases, transcripts of two different sizes were homologous to the cloned sequence. In all four cases, the homologous transcripts appeared at about 7 hr and were decreasing in amount by 13 hr. These results provide evidence for transcriptional control of genes expressed during sporulation and for at least one group of genes that is turned on at about the time of meiosis I in sporulation.
用载体λCharon 28构建的酿酒酵母DNA文库，检测与从产孢良好的酵母菌株AP1 a/α中分离的聚腺苷酸加尾RNA（poly(A)+ RNA）所制备的cDNA互补的序列。RNA是从在产孢培养基中培养7、9、11和13小时的细胞中分离得到的。以与α/α（不产孢）AP1的聚腺苷酸加尾RNA互补的DNA作为对照，在对相当于三个酵母基因组的DNA进行筛选时，获得了46个噬菌体，它们与a/α cDNA的反应比与α/α cDNA的反应更强。其中两个噬菌体似乎含有a/α特异性基因，因为它们与营养期a/α RNA的cDNA杂交。其余的似乎对应于在产孢过程中优先表达的a/α基因。对四个克隆序列进行限制性内切酶分析，结果显示每个序列都有一个单一的主要转录区域；这些区域大小在2.5至4.0千碱基之间。RNA印迹分析表明，在这四个案例中的三个案例中，两种不同大小的转录本与克隆序列同源。在所有四个案例中，同源转录本在大约7小时出现，到13小时时数量减少。这些结果为产孢过程中表达的基因的转录调控以及至少一组在产孢过程中减数分裂I时开启的基因提供了证据。
