Isolation and characterization of sperm associated genes

我们从一个消减的cDNA文库中克隆了单倍体生殖细胞特异的cDNA，该消减文库是从35日龄小鼠的睾丸中减去17日龄小鼠睾丸的mRNA(在单倍体生殖细胞发育之前)而产生的。详细的mRNA表达分析表明，与克隆的cDNA对应的基因在生殖细胞的所有分化阶段、特定的分化阶段或单倍体生殖细胞发育的特定步骤中都有特异的表达。所有这些基因的表达都是发育控制的。生殖细胞特异性基因编码产物的产物包括在细胞内稳态方面起各种作用的蛋白质。特别是，一些蛋白质代表了先前已知的能量代谢酶的生殖细胞特异性同工酶。我们还分离了单倍体特异基因的基因组DNA，并在基因启动子区域确定了一些转录所必需的调控基序。其中一个基序，环状AMP反应元件，存在于几个睾丸特异基因的启动子区域，并被认为具有重要的功能。然而，haspin基因启动子区域不包含Cre基序。同样，基质金属蛋白酶-28的启动子区域(Illman等人)。2001)，激素敏感脂肪酶(Blaise等人。2001)、ldhc(Jethanandani和Goldberg 2001)、SP-10(Reddi等人1999)，在单倍体生殖细胞中特异表达的基因不包含Cre基序。这些发现表明，存在不同的单倍体生殖细胞特异性调控蛋白，它们专门调控单倍体生殖细胞特异性基因的表达。在这里，我们描述了我们关于生殖细胞特异性基因和基因、产物的最新发现，并讨论了我们的方法与生殖细胞分化机制研究的相关性。

We have cloned haploid germ-cell-specific cDNAs from a subtracted cDNA library that was generated by subtracting the mRNA of 17-day-old mouse testes (before haploid germ cells develop) from the cDNA of 35-day-old mouse testes. Detailed mRNA expression analysis revealed that the genes corresponding to the cloned cDNAs were exclusively expressed in germ cells at all steps of differentiation, at specific steps of differentiation, or at specific steps in the development of haploid germ cells. The expression of all of these genes was developmentally controlled. The products of germ-cell-specific genes encoded products included proteins of all sorts of roles in aspect of cellular homeostasis. Particularly, some proteins represented germ-cell-specific isozymes of previously known enzymes in energy metabolism. We also isolated the genomic DNA of haploid-specific genes and identified a number of regulatory motifs in the gene-promoter regions that were essential for transcription. One of these motifs, the cyclic AMP response element, was present in the promoter regions of several testis-specific genes, and was deemed to be functionally important. However, the haspin gene-promoter region did not contain a CRE motif. Similarly, the promoter regions of the MMP-28 (Illman et al. 2001), Hormone-sensitive lipase (Blaise et al. 2001), ldhc(Jethanandani and Goldberg 2001), SP-10(Reddi et al. 1999) genes which were specifically expressed in haploid germ cells, did not contain CRE motifs. These findings suggest the existence of different haploid germ-cell-specific regulatory proteins that specifically regulate the expression of haploid germ-cell-specific genes. Here, we describe our recent findings regarding germ-cell-specific genes and gene, products, and we discuss the relevance of our approaches to the study of germ cell differentiation mechanisms.