文章摘要
翘嘴鲌2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分析
Molecular characterization of two growth hormone receptor (GHR) genes, and association analysis between microsatellite polymorphism and growth traits in the topmouth culter, Culter alburnus
投稿时间:2019-02-22  修订日期:2019-04-11
DOI:
中文关键词: 翘嘴鲌  生长激素受体  序列分析  微卫星  生长性状
英文关键词: Culter alburnus  Growth hormone receptor  Sequence analysis  Microsatellite  Growth trait
基金项目:浙江省农业新品种选育重大科技专项, 2016C02055-1
作者单位E-mail
刘士力 浙江省淡水水产研究所 liushili1212@126.com 
贾永义 浙江省淡水水产研究所 yongyi_jia@163.com 
刘加林 浙江省淡水水产研究所  
郑建波 浙江省淡水水产研究所  
迟美丽 浙江省淡水水产研究所  
程 顺 浙江省淡水水产研究所  
蒋文枰 浙江省淡水水产研究所  
顾志敏 浙江省淡水水产研究所 guzhimin2006@163.com 
赵金良 上海海洋大学  
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中文摘要:
      为了更好地研究翘嘴鲌两种生长激素受体(Growth hormone receptor, GHR)的结构和功能, 本研究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3 498 bp,开放阅读框 (ORF)为1 818 bp,编码605个氨基酸;GHR2的cDNA序列长度为1 743bp,ORF为1 743 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%。两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,这些差异表明两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10kb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属于高度多态位点(PIC>0.5),第6个内含子中的(GAAG)5属中度多态位点(PIC=0.463)。第7内含子中的两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辅助育种提供助力与参考。
英文摘要:
      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 were 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 818bp, and a 605 amino acid residue encoded protein. The full length of GHR2 cDNA is 1 743bp, and with an ORF of 1 743bp, 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 is GHR1 is over 10 kb so that they weren't 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, which had good potential for individual identification. Correlation analysis indicated that the four polymorphic microsatellite loci were all closely related to the growth traits. The cloning and the characterization of microsatellites in GHR gene provide a reference for further study of its biological function and molecular marker assisted breeding in C. alburnus.
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