Analysis of genes which are Involved In regulation for sex differentiation of amphibians

1 ． SF-1/Ad4BP从蛙睾丸λgt10 cDNA文库中成功克隆到SF-1/Ad4BP cDNA。它编码了一个468个氨基酸的蛋白，其ORF与鸡的OR2.1 （SF-1/Ad4BP同源物）有70%的相似性，与牛的Ad4BP有62%的相似性。SF-1/Ad4BP mRNA在成年蛙的睾丸、脑、肾和脾脏均有表达，卵巢无表达。利用该cDNA作为探针，从λ gt10基因组DNA文库中克隆出SF-1/Ad4BP基因的5 '上游区域，包括第1和第2外显子。对该基因的序列分析表明，另一个核因子ELP可能不是由同一基因转录而来。我们克隆了青蛙FTZ-F1 α和rrFTZ-F1 β cdna的同源物。在12期胚胎中表达两种类型（α和β）的rrFTZ-F1 mrna。α和β mrna在25期蛙的睾丸中表达增强，在蜕变2个月后表达最明显。此外，我们克隆了rrFTZ-F1的外显子I或II缺失的区域。基因组结构分析显示，rrFTZ-F1 p是rrFTZ-F1 α.3外显子i部分截断的变体。我们从成年蛙睾丸cdna中分离出两种不同的Sox9基因mRNA亚型（α和β）。Sox9在16期后的胚胎中表达上调，在发育中的睾丸和卵巢中均可见。Northern blot测定Sox9转录本的大小为7.8 knt。此外，Sox9 α在成年蛙的睾丸和大脑各组织中均有显著表达，且显著高于Sox9 β。Sox9在两性中都有表达的事实表明，该基因参与了雄性和雌性青蛙的性腺发育。

1 . SF- I /Ad4BPWe successfully cloned the SF-1/Ad4BP cDNA from the testis λgt10 cDNA library of the frog, Rana rugosa. It encoded a protein of 468 amino acids and its ORF shared 70% similarity with that of chicken OR2.1 (a SF-1/Ad4BP homologue) and 62% with bovine Ad4BP. SF-1/Ad4BP mRNA was expressed in testes, brains, kidneys and spleens , but not ovaries of adult frogs. The 5 ' -upstream region of the SF-1/Ad4BP gene including the 1st and 2nd exons was also cloned from a λ gt10 genomic DNA library using the cDNA as a probe. A sequence analysis of this gene showed that another nuclear factor ELP probably is not transcribed from the same gene.2. FTZ-F 1We cloned frog homologues of FTZ-F1 α and rrFTZ-F1 β cDNAs. Two types (α and β) of rrFTZ-F1 mRNAS were expressed in the embryo at stage 12. Expressions of both the α and β mRNAs became stronger in the testis of frogs at stage xxv and were the most prominent in that of frogs 2 months after metamorphosis. In addition, we cloned the regions with either exon I or II of the rrFTZ-F1 gone. Genomic structure analysis revealed that rrFTZ-F1 p is a partial exon I-truncated variant of rrFTZ-F1 α.3. Sox9We isolated two different mRNA isoforms ( α and β ) of the frog Sox9 gene from adult frog testis cDNAS. Sox9 expression was up-regulated in embryos after stage 16, and was seen in both developing testes and ovaries. The size of Sox9 transcripts was determined to he 7.8 knt by Northern blot analysis. In addition Sox9 α expression was found prominently in the testis and brain among various tissues of adult frogs examined, and was considerably higher than Sox9 β. The fact that Sox9 is expressed in both sexes suggests that this gene is involved in gonadal development of male and female frogs.