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

为了系统地分离控制细胞周期和细胞生长的基因，我们建立了一种改进的方法来制备高质量的消减cDNA文库，从而可以大规模分离转录诱导的mRNA(CDNAs)物种。使用该方法，我们制备了几个实验系统的消减cDNA文库，如下所述。为了对消减文库中涉及的克隆进行有效和全面的分离，我们还开发了一种逐步消减方法。通过应用这些技术，我们在以下实验系统中分离了大量新的cDNA克隆。它们被综合命名为TISP(精子发生中转录增加)、MEU(减数分裂表达上调)、EPOC(主要在破骨细胞中表达)、TIGA(通过生长停滞而上调转录)、ELM(在MI小鼠中表达上调)、TIB(在BL6小鼠中转录上调)、TRIF(在F2408中转录下调)和SSR(剪应力反应)。通过DNA测序，我们发现这些基因中约有一半是新基因，其中一些甚至还没有在基因库中注册为EST(Exposed Sequence Tag)。我们研究了这些基因的功能，发现它们中的许多都具有重要的生物学功能。这些结果表明，我们的策略适用于各种各样的生物现象，其中转录上调或下调在这些现象中起着重要作用，并有助于理解其他无法解决的生物学问题的调控机制。

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.