水产学报2012，Vol.36Issue(8)：1179-1184,6.DOI:10.3724/SP.J.1231.2012.27941
大鲵Dmrt2基因cDNA的克隆与表达分析
Molecular cloning and expression analysis of Dmrt2 gene in Andrias davidianus
摘要
Abstract
Dmrt family is one of the oldest developmental gene family, and is important to the embryo normal growth, spermatogenesis, and the nervous system and sense organ development. Dmrt2 is one of the members of Dmrt family, which was cloned in mammals such as human, birds such as Gallus gallus, aquatic animals such as Oryzias latipes, is closely related to gonad development and growth. RACE -PCR technique was used to clone Dmrti gene in Andrias davidianus, 1 026 bp full-length cDNA was obtained, containing 1 581 bp open reading frame(ORF), 58 bp 5'UTR, 387 bp 3'UTR, GenBank number is FJ859987. This sequence encoded 526 amino acids, and the amino acid sequence was across the membrane protein, no signal peptide, and the protein positioning in the cytoplasm. The amino acid sequence was analyzed by the Conserved Domain Database(CDD), surveyed that single amino-terminal DM domain is from 93 bp to 146 bp, contains a pattern of conserved zinc chelating residues C2H2C4, super-family pfam00751 and smart003010. Blast DM domain with Homo sapiens mab-3, G gallus Dmrt2, Xenopus (Silurana) tropicalis Dmrti and Danio rerio mab-3, five amino acid had variation, include Ile, Arg, Met, Thr, Cys. Phylogenetic analysis indicated that the five species of Homo sapiens , G. gallus , X. (Silurana) tropicalis and D. rerio were clustered first, that means Dmrt family is highly conserved during evolution, and the five amino acids variation had little effect on protein functions. RT-PCR was used to detect the expression of Dmrt2 in skin, intestines, gills, muscle, stomach, spermary and kidney of A. davidianus, and results showed it is high level expression only in spermary and muscle, which indicated that the Dmrti gene may play an important role in gonad development and growth development.关键词
大鲵/Dmrt2/实时荧光定量PCRKey words
Andrias davidianus/ Dmrt2/ real-time PCR分类
生物科学引用本文复制引用
许宝红,肖真明,肖调义,陈开健,刘巧林,周伟,刘敏,姚一彬..大鲵Dmrt2基因cDNA的克隆与表达分析[J].水产学报,2012,36(8):1179-1184,6.基金项目
国家自然科学基金项目(31040083) （31040083）
湖南省科技厅攻关项目(06NK3049) （06NK3049）
湖南省教育厅重点项目(07A012) （07A012）
