花鲈和西伯利亚鲟生长激素/类胰岛素样生长因子-Ⅰ轴相关基因全长cDNA的克隆和序列分析
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摘要
本研究采用反转录聚合酶链式反应(RT-PCR)和cDNA末端快速扩增(RACE)技术克隆花鲈(Lateolabrax japonicus)和西伯利亚鲟(Acipenser baerii Brandt)生长激素(GH)和生长激素受体(GHR)cDNA全长序列。结果显示:花鲈GH(Gen Bank登录号JQ995145)cDNA全长949bp,开放阅读框615 bp,编码204个氨基酸,预测分子质量为23.06 ku,与其他物种的相似性为36.0%~90.2%;花鲈GHRⅠ(Gen Bank登录号JX402001)cDNA全长3 070 bp,开放阅读框1 911 bp,编码637个氨基酸,预测分子质量为70.79 ku,与其他物种的相似性为32.2%~86.2%;花鲈GHRⅡ(Gen Bank登录号JQ995146)cDNA全长2 926 bp,开放阅读框1 749 bp,编码582个氨基酸,预测分子质量为64.42 ku,与其他物种的相似性为30.8%~81.4%。西伯利亚鲟GH(Gen Bank登录号JX003684)cDNA全长999 bp,开放阅读框645 bp,编码214个氨基酸,预测分子质量为24.14 ku,与其他物种的相似性为38.3%~70.1%;西伯利亚鲟GHR(Gen Bank登录号JX003685)cDNA全长2 283 bp,开放阅读框1 716 bp,编码571个氨基酸,预测分子质量为63.41 ku,与其他物种的相似性为39.5%~49.2%。花鲈和西伯利亚鲟GH和GHR cDNA全长序列的获得为进一步研究鱼类GH/类胰岛素样生长因子-Ⅰ(IGF-Ⅰ)轴对生长发育的调控机制奠定了科学基础。
The reverse transcription-phosphatase chain reaction(RT-PCR) and rapid amplification of c DNA ends(RACE) methods were used in this experiment to clone the full-length sequences of c DNAs for growth hormone(GH) and GH receptor(GHR) of Japanese seabass(Lateolabrax japonicus) and Siberian sturgeon(Acipenser baerii Brandt).The results showed as follows: the full-length G H c DNA of Japanese seabass(Gen Bank accession No.JQ995145) was 949 bp,consisting of an open reading frame(ORF) of 615 bp encoding a protein of204 amino acids with a theoretical molecular weight of 23.06 ku,and shared 36.0% to 90.2% similarity with other species.The full-length G HR Ⅰ c DNA of Japanese seabass(Gen Bank accession No.JX402001) was 3070 bp,consisting of an ORF of 1 911 bp encoding a protein of 637 amino acids with a theoretical molecular weight of 70.79 ku,and shared 32.2% to 86.2% similarity with other species.The full-length G HRⅡ c DNA of Japanese seabass(Gen Bank accession No.JQ995146) was 2 926 bp,consisting of an ORF of 1 749 bp encoding a protein of 582 amino acids with a theoretical molecular weight of 64.42 ku,and shared 30.8% to 81.4%similarity with other species.The full-length G H c DNA of Siberian sturgeon(Gen Bank accession No.JX003684) was 999 bp,consisting of an ORF of 645 bp encoding a protein of 214 amino acids with a theoretical molecular weight of 24.14 ku,and shared 38.3% to 70.1% similarity with other species.The full-length G HR c DNA of Siberian sturgeon(Gen Bank accession No.JX003685) was 2 238 bp,consisting of an ORF of 1 716 bp encoding a protein of 571 amino acids with a theoretical molecular weight of 63.41 ku,and shared 39.5% to49.2% with other species.The full-length sequences of c DNAs for G H and G HR of Japanese seabass and Siberian sturgeon will provide a scientific basis for further study on the regulatory mechanism of GH/IGF-Ⅰ axis on growth and development of fish.
The reverse transcription-phosphatase chain reaction(RT-PCR) and rapid amplification of c DNA ends(RACE) methods were used in this experiment to clone the full-length sequences of c DNAs for growth hormone(GH) and GH receptor(GHR) of Japanese seabass(Lateolabrax japonicus) and Siberian sturgeon(Acipenser baerii Brandt).The results showed as follows: the full-length G H c DNA of Japanese seabass(Gen Bank accession No.JQ995145) was 949 bp,consisting of an open reading frame(ORF) of 615 bp encoding a protein of204 amino acids with a theoretical molecular weight of 23.06 ku,and shared 36.0% to 90.2% similarity with other species.The full-length G HR Ⅰ c DNA of Japanese seabass(Gen Bank accession No.JX402001) was 3070 bp,consisting of an ORF of 1 911 bp encoding a protein of 637 amino acids with a theoretical molecular weight of 70.79 ku,and shared 32.2% to 86.2% similarity with other species.The full-length G HRⅡ c DNA of Japanese seabass(Gen Bank accession No.JQ995146) was 2 926 bp,consisting of an ORF of 1 749 bp encoding a protein of 582 amino acids with a theoretical molecular weight of 64.42 ku,and shared 30.8% to 81.4%similarity with other species.The full-length G H c DNA of Siberian sturgeon(Gen Bank accession No.JX003684) was 999 bp,consisting of an ORF of 645 bp encoding a protein of 214 amino acids with a theoretical molecular weight of 24.14 ku,and shared 38.3% to 70.1% similarity with other species.The full-length G HR c DNA of Siberian sturgeon(Gen Bank accession No.JX003685) was 2 238 bp,consisting of an ORF of 1 716 bp encoding a protein of 571 amino acids with a theoretical molecular weight of 63.41 ku,and shared 39.5% to49.2% with other species.The full-length sequences of c DNAs for G H and G HR of Japanese seabass and Siberian sturgeon will provide a scientific basis for further study on the regulatory mechanism of GH/IGF-Ⅰ axis on growth and development of fish.
引文
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[2]张志勇,薛敏,王嘉,等.混合植物蛋白质替代鱼粉对花鲈和西伯利亚鲟生长和肉质影响的比较研究[J].动物营养学报,2013,25(6):1260-1275.
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[7]马细兰,张勇,黄卫人,等.尼罗罗非鱼生长激素及其受体的c DNA克隆与mRNA表达的雌雄差异[J].动物学报,2006,52(5):924-933.
[8]李胜杰,白俊杰,叶星,等.加州鲈生长激素和胰岛素样生长因子I c DNA的克隆及序列分析[J].广东海洋大学学报,2007,27(3):1-5.
[9]GARDINER B G.Sturgeons as living fossils[M]//ELDREDGE N,STANLEY S M.Living fossils.NewYork:Springer Verlag Press,1984:148-152.
[10]CALDUCH-GINER J A,MINGARRO M,DE CELIS S V R,et al.M olecular cloning and characterization of gilthead sea bream(Sparus aurata)growth hormone receptor(GHR).Assessment of alternative splicing[J].Comparative Biochemistry and Physiology Part B:Biochemistry and M olecular Biology,2003,136(1):1-13.
[11]VERY N M,KITTILSON J D,NORBECK L K,et al.Isolation,characterization,and distribution of two c DNAs encoding for growth hormone receptor in rainbowtrout(Oncorhynchus mykiss)[J].Comparative Biochemistry and Physiology Part B:Biochemistry and M olecular Biology,2005,140(4):615-628.
[12]SAERA-VILA A,CALDUCH-GINER J A,PREZSNCHEZ J.Duplication of growth hormone receptor(GHR)in fish genome:gene organization and transcriptional regulation of G HR typeⅠandⅡin gilthead sea bream(Sparus aurata)[J].General and Comparative Endocrinology,2005,142(1/2):193-203.
[13]章力,黄希贵,焦保卫,等.南方鲇两种生长激素受体的结构分析及其组织分布和激素调节[J].动物学报,2006,52(6):1096-1106.
[2]张志勇,薛敏,王嘉,等.混合植物蛋白质替代鱼粉对花鲈和西伯利亚鲟生长和肉质影响的比较研究[J].动物营养学报,2013,25(6):1260-1275.
[3]ROSE T M,SCHULTZ E R,HENIKOFF J G,et al.Consensus-degenerate hybrid oligonucleotide primers for amplification of distantly related sequences[J].Nucleic Acids Research,1998,26(7):1628-1635.
[4]KUMAR S,TAMURA K,JAKOBSEN I B,et al.M EGA2:molecular evolutionary genetics analysis software[J].Bioinformatics,2001,17(12):1244-1245.
[5]PREZ-SNCHEZ J,LE BAIL P Y.Growth hormone axis as marker of nutritional status and growth performance in fish[J].Aquaculture,1999,177(1/2/3/4):117-128.
[6]AYSON F G,DE JESUS E G T,AMEMIYA Y,et al.Isolation,c DNA cloning,and growth promoting activity of rabbitfish(Siganus guttatus)growth hormone[J].General and Comparative Endocrinology,2000,117(2):251-259.
[7]马细兰,张勇,黄卫人,等.尼罗罗非鱼生长激素及其受体的c DNA克隆与mRNA表达的雌雄差异[J].动物学报,2006,52(5):924-933.
[8]李胜杰,白俊杰,叶星,等.加州鲈生长激素和胰岛素样生长因子I c DNA的克隆及序列分析[J].广东海洋大学学报,2007,27(3):1-5.
[9]GARDINER B G.Sturgeons as living fossils[M]//ELDREDGE N,STANLEY S M.Living fossils.NewYork:Springer Verlag Press,1984:148-152.
[10]CALDUCH-GINER J A,MINGARRO M,DE CELIS S V R,et al.M olecular cloning and characterization of gilthead sea bream(Sparus aurata)growth hormone receptor(GHR).Assessment of alternative splicing[J].Comparative Biochemistry and Physiology Part B:Biochemistry and M olecular Biology,2003,136(1):1-13.
[11]VERY N M,KITTILSON J D,NORBECK L K,et al.Isolation,characterization,and distribution of two c DNAs encoding for growth hormone receptor in rainbowtrout(Oncorhynchus mykiss)[J].Comparative Biochemistry and Physiology Part B:Biochemistry and M olecular Biology,2005,140(4):615-628.
[12]SAERA-VILA A,CALDUCH-GINER J A,PREZSNCHEZ J.Duplication of growth hormone receptor(GHR)in fish genome:gene organization and transcriptional regulation of G HR typeⅠandⅡin gilthead sea bream(Sparus aurata)[J].General and Comparative Endocrinology,2005,142(1/2):193-203.
[13]章力,黄希贵,焦保卫,等.南方鲇两种生长激素受体的结构分析及其组织分布和激素调节[J].动物学报,2006,52(6):1096-1106.