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

1 .一、SF-1/Ad 4 BP我们从中国林蛙（Rana rugosa）睾丸λ gt 10 cDNA文库中成功克隆了SF-1/Ad 4 BP cDNA。该基因编码468个氨基酸，其ORF与鸡OR 2.1（SF-1/Ad 4 BP同源物）的相似性为70%，与牛Ad 4 BP的相似性为62%。SF-1/Ad 4 BPmRNA在成年蛙的睾丸、脑、肾和脾中均有表达，但在卵巢中不表达。以该cDNA为探针，从λ gt 10基因组DNA文库中克隆了SF-1/Ad 4 BP基因5 '上游区，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的. FTZ-F 1克隆了青蛙FTZ-F 1 α和rrFTZ-F 1 β的同源基因。两种类型（α和β）的rrFTZ-F1 mRNAS在第12阶段的胚胎中表达。α和β mRNA在xxv期的精巢中表达增强，在变态后2个月的精巢中表达最强。此外，我们克隆了rrFTZ-F1外显子I或II缺失的区域。基因组结构分析表明，rrFTZ-F1 p是rrFTZ-F1 α.3的部分外显子I截短变体。Sox 9我们从成年青蛙睾丸cDNA中分离到两种不同的青蛙Sox 9基因mRNA亚型（α和β）。Sox 9表达在16期后的胚胎中上调，并且在发育中的睾丸和卵巢中均可见。通过北方印迹分析确定Sox 9转录物的大小为7.8 knt。此外，在所检测的成年蛙的各种组织中，Sox 9 α在睾丸和脑中的表达显著高于Sox 9 β。Sox 9在两种性别中表达的事实表明，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.