黄颡鱼Pelteobagrus fulvidraco AMH基因的克隆鉴定及表达

Cloning Identification and Expression Analysis of AMH Gene in Yellow Catfish(Pelteobagrus fulvidraco)

为研究抗缪勒氏管激素AMH对黄颡鱼Pelteobagrus fulvidraco性别分化及生长发育的调控作用,克隆扩增了黄颡鱼AMH基因的全长序列(GenBank检索号MK310106),研究该基因的组织表达模式。结果表明:AMH基因的全长为3588bp,其开放阅读序列为1653bp,编码550个氨基酸,位于第2号染色体上(19371164~19374761bp)。同源性分析表明,与已发布的黄颡鱼Tachysurus fulvidraco的相似性最高,可达99.6%;与低眼无齿[鱼芒]Pangasianodon hypophthalmus及斑点叉尾鮰Ictalurus punctatus的相似性也很高,分别达75.8%与74.8%,而与哺乳动物相似性较低。系统进化树分析显示:黄颡鱼AMH可与斑点叉尾鮰、低眼无齿[鱼芒]等鲶形目物种聚集成簇。Real-TimePCR检测结果表明:AMH的mRNA在各组织中的表达量差异性极大,在性腺组织中表达量最高,其次是肝脏和脑;1龄精巢的表达量最高,2龄次之,而1、2龄雌鱼的表达量基本相近。AMH对黄颡鱼性腺的分化及生长发育有重要调控作用。

Full-length cDNA sequence of AMH was cloned and amplified in yellow catfish Pelteobagrus fulvidraco (GenBank accession number MK310106), and its expression pattern was investigated to evaluate the role of antik-Müllerian hormone (AMH) in sex differentiation, growth, and development of yellow catfish. It was found that there was 3 588 bp in full-length of AMH gene, with open reading frame (ORF) of 1 653 bp encoding 550 amino acids, being located on site 19 371 164 ~ 19 374 761 bp of chromosome 2. AMH of yellow catfish was shared 75.8% of similarity with Pangasianodon hypophthalmus and 74.8% of similarity with channel catfish Ictalurus punctatusin sequence, with the maximal similarity (99.6%) in sequence with that of Tachysurus fulvidraco and the minimal similarity with mammal. Phylogenetic tree revealed that AMH of yellow catfish was clustered with Siluriformes including channel catfish and Pangasianodon hypophthalmus. Real-time PCR showed that the maximal mRNA expression of AMH was found in gonad, followed by liver and brain. The maximal expression of gonad was observed in 1-year-old males, followed by 2-year-old males, and similar in 1-year-old and 2-year-old females. The findings indicated that AMH played an important role in regulation of sex differentiation, growth and development of yellow catfish.