草鱼PAX7基因的克隆、序列分析及组织表达  

Cloning and Tissue Expression Analysis of PAX7 Gene in Grass Carp(Ctenopharyngodon idellus)

作　　者：甘甜[1] 冷向军[1] 郭婷[1] 胡静[1] 魏静[1] 李小勤[1] 

机构地区：[1]上海海洋大学水产与生命学院教育部水产种质资源发掘与利用重点实验室,上海201306

出　　处：《生物技术通报》2014年第11期152-158,共7页Biotechnology Bulletin

基　　金：上海市自然科学基金项目(13ZR14 19600)

摘　　要：配对盒基因7(Paired box 7 gene,PAX7)在神经嵴发育及原肠胚形成、肌肉自主更新与再生中扮演着重要角色,对细胞更新、分化、凋亡等起着十分重要的调控作用。参照Gen Bank中斑马鱼、线鳍电鳗和虹鳟等物种PAX7序列的保守区域设计简并引物,进行反转录PCR扩增,获得草鱼PAX7基因的部分序列,长645 bp,编码214个氨基酸,包含编码128个氨基酸的PAX7蛋白配对框(PD)。氨基酸序列比对结果发现,草鱼PAX7基因与其他物种具有高度的相似性,与斑马鱼、线鳍电鳗、日本青鳉、金头鲷、虹鳟、大西洋鲑、北极嘉鱼、人、野牦牛、褐家鼠和小鼠的PAX7氨基酸序列同源性可达90%-97%;各物种PAX7部分氨基酸序列的系统进化树结果显示,草鱼与斑马鱼、日本青鳉、北极嘉鱼、大西洋鲑、虹鳟、线鳍电鳗和金头鲷聚为一大支,小鼠、褐家鼠,野牦牛与人类另聚一支,所得结果符合物种传统分类进化地位。运用实时荧光定量PCR检测草鱼PAX7基因在各组织中的差异表达,结果显示PAX7基因在肌肉中表达量最高;其次是前肠和皮肤,在心、脑、肾和肝中仅少量表达,所得结果与该基因的功能相一致。Paired box gene 7（PAX7）is crucially important to the cellular renewal, differentiation and apoptosis, especially in neural crest development, gastrulation, and muscle self-renewal. ThePAX7 domains sequence is conserved among several species, such as zebrafish, apteronotidae, and rainbow trout, and the conserved sequence zebrafish was used to design degenerate primers for reverse-transcription PCR（RT-PCR）. A partial sequence ofPAX7 from grass carp was obtained for a 645 bp segment encoding a 214 amino acid peptide, containing a paired box domain with 128 amino acids. The deduced protein showed homology to zebrafish（Danio rerio）, apteronotidae（Sternopygus macrurus）, Japanese Medaka（Oryzias latipes）, gilthead bream（Sparus aurata）, rainbow trout（Oncorhynchus mykiss）, Atlantic salmon（Salmo salar）, Arctic charr（Salvelinus alpinus）, human（Homo sapiens）, wild yak（Bos grunniens mutus）, brown rat（Rattus norvegicus）, and mouse（Mus musculus）with 90%-97% identities. Analysis of the PAX7 phylogenetic tree revealed that the grass carp joined with zebrafish, and there was a confluence of Japanese medaka, Arctic charr, Atlantic salmon, rainbow trout, apteronotidae and gilthead bream. The other branch was consisted of mouse, brown rat, wild yak and human. These results conformed to the traditional species classification evolution status. The expression in tissues was detected by Real-time quantitative PCR, which indicated that the highest level ofPAX7 gene expression was found in muscle, a lower expression level was found in foregut and skin, and the lowest detectable level was found in heart, brain, kidney and liver. The results are in agreement with the function of the gene.

关 键 词：PAX7基因 草鱼 序列分析 组织差异表达 

分 类 号：S917.4[农业科学—水产科学]