小体鲟卵透明带家族ZP3亚型和ZPAX基因cDNAs克隆及其组织表达分析  

cDNAs cloning and expression of egg zona pellucida ZP3s and ZPAX genes in various tissues of sterlet Acipenser ruthenus

作　　者：东天[1] 朱华[1] 王巍[1] 董颖[1] 田照辉[1] 胡红霞[1] DONG Tian;ZHU Hua;WANG Wei;DONG Ying;TIAN Zhao-hui;HU Hong-xia(Beijing Key Laboratory of Fishery Biotechnology,Beijing Fisheries Research Institute and National Engineering Research Centre for Freshwater Fisheries,Beijing 100068,China)

机构地区：[1]北京市水产科学研究所渔业生物技术北京市重点实验室,北京100068

出　　处：《大连海洋大学学报》2018年第4期444-454,共11页Journal of Dalian Ocean University

基　　金：国家科技支撑计划项目(2015BAD25B00);中央引导地方科技发展专项(Z161100004516003);北京市鲟鱼;鲑鳟鱼创新团队项目(SCGWZJ20171102-1)

摘　　要：为研究透明带ZP家族在小体鲟Acipenser ruthenus性腺发育、卵子发生等过程中所发挥的作用,采用PCR技术克隆获得小体鲟卵透明带家族ZP3 3种亚型(ArZP3-1、ArZP3-4、ArZP3-5)及ZPAX基因(ArZPAX)cDNA部分序列。结果表明:ArZP3-1部分cDNA序列长度为1029 bp,对应编码342个氨基酸;ArZP3-4部分cDNA序列长度为1191 bp,对应编码397个氨基酸;ArZP3-5部分cDNA序列长度为717 bp,对应编码238个氨基酸;ArZPAX部分cDNA序列长度为733 bp,对应编码243个氨基酸;ArZP3蛋白3种亚型及ArZPAX蛋白氨基酸序列中均含有一个保守的ZP结构域;基于ZP氨基酸序列的系统发育树显示,小体鲟ArZP3蛋白3种亚型与高等鱼类ZP3亲缘关系较近,而ArZPAX蛋白则与哺乳类、鸟类、高等鱼类和爬行类ZPAX亲缘关系均较远;通过半定量RT-PCR进行组织表达特异性分析表明,ArZP3-1基因在小体鲟卵巢、精巢、肝、脑、肾、心、肌肉、鳃中表达,ArZP3-4基因仅在小体鲟卵巢和精巢中表达,而ArZPAX基因在小体鲟卵巢、精巢、肝、肠、肾、脾、心、鳃、脑中均有表达,3个基因在性腺中的表达量均最高,且均具有雌、雄差异性表达特点。The partial-length cDNA sequences of ArZP3 s(ArZP3-1,ArZP3-4 and ArZP3-5)and ArZPAX genes are cloned in sterlet Acipenser ruthenus by PCR to identify the gonadal development mechanism of zona pellucida gene family in sterlet.It was found that ArZP3-1 was 1029 base pairs(bp),predicting proteins of 342 amino acid residues;ArZP3-4 1191 bp in length,predicting proteins of 397 amino acid residues;ArZP3-5 717 bp in length,predicting proteins of 238 amino acid residues;and ArZPAX 733 bp in length,predicting proteins of 243 amino acid residues.Sequence analysis revealed that these genes were all comprised of a conserved ZP domain.Phylogenetic tree indicated that three ArZP3 s of sterlet showed a closer genetic relationship with higher fishes,and ArZPAX was clustered with its counterparts in other vertebrates.Semiquantitative RT-PCR demonstrated that the ArZP3-1 was expressed in ovary,testis,liver,brain,kidney,heart,white muscle and gill,ArZP3-4 only in ovary and testis,and ArZPAX was detected in ovary,testis,liver,intestine,kidney,spleen,heart,brain and gill,with the maximal expression level in the gonads and differentially sexual expressed.

关 键 词：小体鲟 透明带蛋白 ZPAX 基因克隆 组织表达 

分 类 号：S917.4[农业科学—水产科学]