翘嘴鲌两种生长激素受体基因结构及微卫星多态性与生长性状的相关性  

Molecular characterization of two growth hormone receptor genes, and association analysis between microsatellite polymorphism and growth traits in the topmouth culter(Culter alburnus)

作　　者：刘士力[1,2,3] 贾永义[1] 刘加林 郑建波 迟美丽[1] 程顺[1] 蒋文枰[1] 顾志敏[1] 赵金良[2,3] LIU Shili;JIA Yongyi;LIU Jialin;ZHENG Jianbo;CHI Meili;CHENG Shun;JIANG Wenping;GU Zhimin;ZHAO Jinliang(Key Laboratory of Freshwater Aquatic Animal Genetic and Breeding of Zhejiang Province,Key Laboratory of Healthy Freshwater Aquaculture,Ministry of Agriculture and Rural Affairs,Zhejiang Institute of Freshwater Fisheries,Huzhou 313001,China;Laboratory of Freshwater Fisheries Germplasm Resources,Ministry of Agriculture and Rural Affairs,Shanghai Ocean University,Shanghai 201306,China;Shanghai Engineering Research Center of Aquaculture,Shanghai Ocean University,Shanghai 201306,China;School of Life Science,Nanjing Normal University,Nanjing 210000,China)

机构地区：[1]浙江省淡水水产研究所,农业农村部淡水渔业健康养殖重点实验室,浙江省淡水水产遗传育种重点实验室,浙江湖州313001 [2]上海海洋大学,农业农村部淡水水产种质资源重点实验室,上海201306 [3]上海海洋大学,上海水产养殖工程技术研究中心,上海201306 [4]南京师范大学生命科学学院,江苏南京210000

出　　处：《水产学报》2020年第6期894-906,共13页Journal of Fisheries of China

基　　金：浙江省农业新品种选育重大科技专项(2016C02055-1)。

摘　　要：为了更好地研究翘嘴鲌两种生长激素受体(GHR)的结构和功能,实验以翘嘴鲌转录组中获得的mRNA为基础,对其DNA序列进行了克隆。在进行生物信息学分析的同时,对其中的多态性微卫星位点在120尾同批繁殖、同塘养殖的翘嘴鲌个体中进行了分析。GHR1的cDNA序列长度为3498 bp,开放阅读框(ORF)为1818 bp,编码605个氨基酸;GHR2的cDNA序列长度为1743 bp,ORF为1743 bp,编码580个氨基酸;GHR1和GHR2氨基酸序列均由信号肽、胞外区、跨膜区、胞内区组成,相似度为37.2%。二者在结构上存在较大的差异:GHR1胞外区有7个半胱氨酸残基,而GHR2只有5个,且GHR1比GHR2多3个N-糖基化位点;在胞内区,GHR1存在10个酪氨酸残基而GHR2只有5个,这些差异表明二者可能具有不同的生物学功能。同源氨基酸序列比对发现,GHR与其他鲤科鱼类的同源基因保守性较高。翘嘴鲌2个GHR各包含9个内含子,其中GHR1内含子1和2序列在10 kb以上,本实验没有对其进行扩增。所获得的序列中共发现了6个微卫星位点:GHR1中微卫星位点(CT)6位于第2外显子中,为信号肽编码序列的一部分,位于第8内含子中的(AC)5经检测没有多态性;GHR2中具有4个微卫星位点,位于第1内含子中的(TG)5及第7个内含子中的(TATC)5(AT)15(AC)11(AT)14(TG)6和(TA)15属于高度多态性位点(PIC>0.5),第6个内含子中的(GAAG)5属中度多态性位点(PIC=0.463)。第7内含子中的2个微卫星位点检测到基因型数目分别为50和61,具有良好的个体识别潜力。关联分析结果表明这4个多态性微卫星位点与生长性状具有一定相关性。翘嘴鲌GHR基因的克隆以及序列中微卫星的特征分析为深入研究其生物学功能及分子标记辅助育种提供助力与参考。To better study the structure and function of the growth hormone receptor(GHR) of the topmouth culter,Culter alburnus, the DNA sequences of GHR1 and GHR2 were cloned based on mRNA data from the transcriptome of C. alburnus. Bioinformatics analysis was performed and the polymorphic microsatellite loci in the GHRs were tested in 120 samples which were bred in the same batch and cultured in the same pond. The full length of GHR1 cDNA is 3 498 bp, with an open reading frame(ORF) of 1 818 bp, and a 605 amino acid residue encoded protein. The full length of GHR2 cDNA is 1 743 bp, and with an ORF of 1 743 bp, and a 580 amino acid residue encoded protein, the amino acid sequences of GHR1 and GHR2 both comprised a signal peptide, extracellular region, transmembrane region, and intracellular region, and are 37.2% similar. There were marked differences in their structures. GHR1 has seven cysteine residues in its extracellular region of GHR1, but GHR2 has only five.GHR1 has three N-glycosylation sites more than GHR2. In the intracellular domain, there are 10 tyrosine residues in GHR1, but only 5 in GHR2, indicating that the two proteins may have different biological functions. Homologous amino acid sequence alignment showed that the GHRs are highly conserved with GHRs from other Cyprinidae. There were both 9 introns in the GHRs of C. alburnus, the length of intron 1 and 2 in GHR1 is over 10 kb so that they were not amplified in this experiment. Six microsatellite loci were found in the obtained sequence: the microsatellite locus(CT)6 in the exon 2 of GHR1 was located in the signal sequence coding region, and no polymorphism of the(AC)5 in intron 8 was detected;there were four microsatellite loci in GHR2, including(TG)5 in intron 1,(TATC)5(AT)15(AC)11(AT)14(TG)6 and (TA)15 in intron 7, which belonged to highly polymorphic loci(PIC > 0.5). The(GAAG)5 microsatellite loci in intron 6 was moderately polymorphic(PIC = 0.463). The number of genotypes detected using two microsatellite loci in intron 7 was 50 and 61, respectively, wh

关 键 词：翘嘴鲌 生长激素受体 序列分析 微卫星 生长性状 

分 类 号：Q786[生物学—分子生物学] S917.4[农业科学—水产科学]