Applied research for the development of gene diagnosis of ovarian dysfunction using a novel ovarian transcription factor GIOT.

为了利用一种新的卵巢转录因子giot建立卵巢功能障碍的基因诊断系统，我们克隆了人giot的同源物，并研究了giot在卵巢功能中的可能作用。构建了含GIOT基因的表达载体，并将其导入大鼠颗粒细胞。同时将含有卵巢基因的报告载体导入细胞内，并检测其活性。转基因研究表明，GIOT可以抑制卵巢基因的表达，并作为卵巢转录抑制因子发挥作用。将该GIOT表达载体导入MA10小鼠LeyDig肿瘤细胞，也获得了类似的结果。因此，我们证明了GIOT实际上对卵巢基因的转录调控起作用。我们还克隆了GIOT的人类同源物。至少分离出了三个人类giot同源物。对这些克隆的核苷酸序列分析表明，这些都是不同的基因。对这些基因的功能分析目前正在进行中。

In order to develop a gene diagnostic system for ovarian dysfunction using a novel ovarian transcription factor GIOT, we have cloned human homologues of GIOT and investigated the possible roles of GIOT for the ovarian functions. An expression plasmid containing GIOT was constructed and rat granulosa cells were transfected with the plasmid. Reporter plasmids containing ovarian genes were simultaneously transfected and the reporter activity was measured. The transfection study demonstrated that GIOT can suppress the ovarian gene expression and works as an ovarian transcription repressor. MA10 mouse Leydig tumor cells were also transfected with the GIOT expression plasmid and similar results were obtained. Therefore, we demonstrated that GIOT actually works on the transcriptional regulation of ovarian genes. We also have cloned human homologues of GIOT. At least three human GIOT homologues were isolated. Nucleotide sequence analysis of these clones revealed that these are distinct genes. Functional analysis of these genes is now underway.