title: Use of stepwise subtraction to comprehensively isolate mouse genes whose transcription is up-regulated during spermiogenesis.
abstract: We report the isolation of 153 mouse genes whose expression is dramatically up-regulated during spermiogenesis. We used a novel variation of the subtractive hybridization technique called stepwise subtraction, wherein the subtraction process is systematically repeated in a stepwise manner. We named the genes thus identified as TISP genes (transcript induced in spermiogenesis). The transcription of 80 of these TISP genes is almost completely specific to the testis. This transcription is abruptly turned on after 17 days of age, when the mice enter puberty and spermiogenesis is initiated. Considering that the most advanced cells present at these stages of spermatogenesis are the spermatids, it is likely that we could isolate most of the spermatid-specific genes. DNA sequencing revealed that about half the TISP genes are novel and uncharacterized genes, confirming the utility of the stepwise subtraction approach for gene discovery.
authors.affiliation: Departments of Molecular Genetics, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Fujii
authors.forename: Kazutoshi
authors.initials: K
authors.lastname: Tamura
authors.forename: Kumiko
authors.initials: K
authors.lastname: Masai
authors.forename: Hiromitsu
authors.initials: H
authors.lastname: Tanaka
authors.forename: Yoshitake
authors.initials: Y
authors.lastname: Nishimune
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nojima
mesh_terms: D000818:Animals; D015152:Blotting, Northern; D018076:DNA, Complementary; D015723:Gene Library; D008297:Male; D051379:Mice; D009693:Nucleic Acid Hybridization; D009928:Organ Specificity; D013091:Spermatogenesis / Q000235:genetics* / Q000502:physiology; D014158:Transcription, Genetic; D015854:Up-Regulation
chemical_list: D018076:DNA, Complementary