The Use of cDNA Libraries to Demonstrate a Linkage Between Transcription and Translation in Male Germ Cells

TL;DRAbstract

cDNA libraries have played a prominent role in developing the extensive database of gene expression in germ cells and somatic cells of the mammalian testis. Differential screening of cDNA libraries has allowed investigators to determine the temporal up-and downregulation of many genes. This chapter discusses how suppressive subtraction hybridization and cDNA sequencing have been used to define populations of messenger RNAs (mRNAs) that selectively bind, or do not bind, to the germ cell-specific Y-box protein, MSY2. MSY2 is an abundant DNA/RNA-binding protein that in vitro binds to all mRNAs, but shows selective binding to a subset of male germ cell mRNAs in cells. This specificity is regulated by MSY2 binding to a conserved sequence in gene promoters, which facilitates MSY2 binding to the transcripts from these promoters in the nucleus and coordinates the transport, storage, and translational suppression of these mRNAs in the cytoplasm.

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