碩士 / 國立屏東科技大學 / 生物科技系所 / 101 / Anti-Mullerian hormone (AMH), also known as Mullerian inhibiting substance (MIS), is a glycoprotein belonging to the transforming growth factor-β superfamily (TGF-β). Reports have shown that teleost AMH is expressed predominantly in the reproductive tissues and acts an important role in gonad development and sex differentiation. In recently years, although the medium-sized fluorescent angelfish (P. scalare) has been established, it is still refused to commercialize to date due to the biosafety concerns. In this regard, infertility control for transgenic fish is a core technology to solve this issue. In this study, AMH cDNA and AMH promoter were cloned from P. scalare. We expected the ablation of gonad can lead to infertility by expressing inducible toxin protein KillerRed using AMH promoter. In present study, a 1545 bp coding region of angelfish AMH cDNA which encoded 514 amino acids was cloned from gonad tissue. Alignment analysis of the AMH with homologs from other fish species showed that AMH consist of a AMH-N domain with 348 amino acids and a TFG-β domain with 93 amino acids, and two consecutive putative plasmin cleavage sites (R/K-XX-R/K) between the AMH-N domain and TFG-β domain. The nucleotide sequences of angelfish (P. scalare) AMH was highly similar to that in tilapla (O. aureus) (82% identical). Tissue distribution showed that AMH gene was specifically expressed in testes and ovary and the expression level of AMH in testes was significantly higher than in ovary. In order to understand the regulatory mechanism, a 5'-upstream region of the AMH gene (2.8 kb) was cloned by genome walking. Putative binding sites analysis for transcription factor binding sites such as SRY, SF-1, Sox 9 and GATA-4 existed within the promoter region. The functional analysis of angelfish (P. scalare) AMH promoter was carried out in zebrafish by expressing a KillerRed reporter gene. Results revealed that the KillerRed was specific-expressed in zebrafish gonad at 25 dpf suggesting angelfish (P. scalare) AMH promoter with tissue specificity.
| Identifer | oai:union.ndltd.org:TW/101NPUS5111010 |
| Date | January 2012 |
| Creators | Jung-Kuang Wang, 汪榮光 |
| Contributors | Shao-Yang Hu, 胡紹揚 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 72 |
Page generated in 0.0015 seconds