Systematic isolation and analysis of novel genes that control cell cycle and cell growth.

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In order to isolate systematically the genes that control cell cycle and cell growth, we have established an improved protocol to prepare a subtracted cDNA library of high quality that allowed the large-scale isolation of transcriptionally induced mRNA (cDNA) species. Using this protocol, we prepared subtracted cDNA libraries of several experimental systems as described below. To perform the efficient and comprehensive isolation of the clones involved in the subtracted cDNA library, we also developed a stepwise subtraction method. By applying these techniques, we have isolated a large number of novel cDNA clones in the following experimental systems. They are comprehensively named as TISP (Transcription increased in spermiogenesis), meu (meiotic expression upregulated), EPOC (Expressed predominantly in osteoclast), TIGA (Transcription increased by growth arrest), ElM (Epxpression increased in mi mouse), TIB (Transcription increased in BL6 mouse), TRIF (Transcription reduced in F2408) and SSR (shear Stress Responsive). By DNA sequencing, we found that about half of these genes are novel, and some of them have not been registered in the gene bank even as EST (expressed sequence tag). We have examined the functions of these genes and found that many of them displayed biologically important functions. These results indicate that our strategy is applicable to a wide variety of biological phenomena in which the trancriptional up or down regulation play the important role in these phenomena and is useful to understand the regulatory mechanisms of otherwise unresolvable biological problems.