Isolation of genes expressed preferentially during sporulation in the yeast Saccharomyces cerevisiae

Abstract

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.