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Cloning and characterization of mouse gelell-specific genes

小鼠 gelell 特异性基因的克隆和表征

基本信息

批准号：
11234202
负责人：
NISHIMUNE Yoshitake
金额：
$ 55.23万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research on Priority Areas
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2002
项目状态：
已结题

项目摘要

Spermiogenesis involves nuclear condensation, the elimination of most of the spermatid cytoplasm, and the formation of the tail and acrosome. We cloned haploid-specific cDNAs from a subtracted cDNA library that was generated by subtracting the mRNA of 17-day-old mouse testis from the cDNA of 35-day-old mouse testis. In this phase, more haploid-cell-specific genes are actively transcribed than was expected ; the expression pattern of each gene with various putative functions is strictly regulated. Elucidating the roles of haploid-germ-cell-specific genes will facilitate understanding of not only spermiogenesis but also of the differences between haploid and diploid cells. To identify the regulatory elements in haploid-specific genes, we isolated genomic DNA. One motif, the cyclic AMP response element (CRE), was present in the promoter regions of various specific genes and is functionally important, as reported previously. However, other genes did not contain CRE motifs. This suggests th … More e existence of different haploid-specific regulatory proteins that regulate the specific expression of these genes. Since many haploid-specific genes are CpG rich, epigenetic regulation via methylation might also contribute to the regulation of their expression. An interesting feature of haploid-specific genes is that a surprisingly high frequency lack introns. The intronless genes may be produced by specific mechanisms that exist in germ cells.More than one tenth of human couples suffer from infertility, and half of these cases are attributable to deficient spermatogenesis in males. Although the molecular basis of most disorders of human spermatogenesis remains unclear, we hypothesized that deterioration in the effect of male germ-cell-specific genes gives rise to male infertility. Our new strategy is to compare the DNA sequences of fertile and infertile men. Since some germ cell-specific genes are intronless, it is easier to examine SNPs or detect mutations by direct DNA sequencing of the PCR-amplified products of chromosomal DNA from blood samples. Less

精子发生包括核浓缩、大部分精子细胞质的消除、尾部和顶体的形成。我们从35日龄小鼠睾丸的cDNA中减去17日龄小鼠睾丸的mRNA而产生的消减cDNA文库中克隆了单倍体特异性cDNA。在这一阶段，更多的单倍体细胞特异性基因被积极转录比预期的，每个基因的表达模式与各种推定的功能受到严格的监管。阐明单倍体生殖细胞特异性基因的作用不仅有助于理解精子发生，而且有助于理解单倍体和二倍体细胞之间的差异。为了鉴定单倍体特异性基因中的调控元件，我们分离了基因组DNA。一个基序，环AMP反应元件（CRE），存在于各种特定基因的启动子区，是功能上重要的，如前所述。然而，其他基因不包含CRE基序。这表明， ...更多信息存在调节这些基因的特异性表达的不同的单倍体特异性调节蛋白。由于许多单倍体特异性基因富含CpG，因此通过甲基化的表观遗传调控也可能有助于其表达的调控。单倍体特异性基因的一个有趣的特征是，令人惊讶的高频率缺乏内含子。无内含子基因可能由存在于生殖细胞中的特定机制产生。超过十分之一的人类夫妇患有不育症，其中一半可归因于男性精子生成不足。虽然大多数人类精子发生障碍的分子基础尚不清楚，但我们假设男性生殖细胞特异性基因的作用恶化会导致男性不育。我们的新策略是比较生育和不育男性的DNA序列。由于一些生殖细胞特异性基因是无内含子的，因此通过对来自血液样品的染色体DNA的PCR扩增产物进行直接DNA测序来检查SNP或检测突变更容易。少