白斑狗鱼Wnt4基因的克隆及其表达分析  

Cloning and Sequence Analysis of Wnt4 Gene in Northern Pike Esox lucius

作　　者：李菁 赵瑞阳 张俊杰[1] LI Jing;ZHAO Ruiyang;ZHANG Junjie(College of Life Science,Xinjiang Agricultural University,Urumqi 830052,China)

机构地区：[1]新疆农业大学生命科学学院,新疆乌鲁木齐830052

出　　处：《水产科学》2023年第3期432-439,共8页Fisheries Science

基　　金：国家自然科学基金资助项目(31660739);新疆维吾尔自治区高校科研计划项目(XJEDU2016I201);新疆农业大学大学生创新项目(201910758040).

摘　　要：为探讨无翅型MMTV整合位点家族成员(Wnt4)基因在白斑狗鱼性别发育过程中的作用,通过反转录PCR法克隆获得白斑狗鱼Wnt4基因片段,运用ClustalX和MEGA6.1软件对其氨基酸同源性及物种进化关系进行分析。研究结果显示,克隆获得的Wnt4基因片段长度为1505bp,其中开放阅读框长度为1059bp,编码352个氨基酸。氨基酸同源性分析表明,白斑狗鱼与虹鳟、青鳉同源性较高,与非洲爪蟾和褐家鼠等相似性则较低。系统发育结果显示,白斑狗鱼与白边锯鳞鱼亲缘关系最近,与海胆和栉孔扇贝、厚壳贻贝的亲缘关系最远。实时荧光定量PCR分析显示,Wnt4基因在白斑狗鱼卵巢、精巢、肠道、肌肉、鳃、脑等组织中均有表达,卵巢的表达最明显,卵巢表达量明显高于精巢,Wnt4基因在卵巢后期发育过程中,表达量呈现下降趋势。研究结果表明,Wnt4基因在白斑狗鱼卵巢发育的早期参与到某些器官的形成及功能的维持。本研究结果可为白斑狗鱼的性别发育机制提供参考资料。The purpose of this study was to investigate the role of Wnt4(wingless-type MMTV integration site family,member 4)gene in sex development of northern pike Esox lucius.The Wnt4 gene fragment was cloned by RT-PCR,the homology of amino acids and the evolutionary relationship of species were analyzed by Clustal X and MEGA 6.1 software.The length of Wnt4 gene fragment was 1505 bp,with the open reading frame of 1059 bp,encoding 352 amino acids.Amino acid homology analysis showed that northern pike had high homology with rainbow trout Oncorhynchus mykiss and medaka Oryzias latipes,but low similarity with Xenopus laevis and Rattus norvegicus.Phylogenetic analysis showed that the phylogenetic relationship between northern pike and Myripristis murdjan was the hithermost,and the phylogenetic relationship between northern pike and Paracentrotus lividus,Azumapecten farreri and Mytilus coruscus was the farthest.Real time fluorescent quantitative PCR analysis showed that Wnt4 gene was expressed in ovary,testis,intestine,muscle,gill and brain.The expression of Wnt4 gene in ovary was the highest,and the expression of Wnt4 gene in ovary was significantly higher than that in testis.With the development of ovary,the expression of Wnt4 gene was decreased.Wnt4 gene was involved in the formation and maintenance of some organs in the early stage of ovarian development.The findings provide reference for the sex development mechanism of northern pike.

关 键 词：白斑狗鱼 Wnt4基因 克隆 生物信息学 组织表达 

分 类 号：Q78[生物学—分子生物学]