建鲤催乳素基因全长cDNA的克隆及序列分析和组织表达
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中央级公益性科研院所基本科研业务费专项基金 (2011JBFC08);现代农业产业技术体系建设专项资金(CARS-46)


Full-length cDNA cloning and tissue expression of PRL from Cyprinus carpio var.Jian
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    摘要:

    为了解催乳素(prolactin,PRL)的基因序列以及在建鲤渗透调节组织中的表达情况,采用同源克隆和末端快速扩增(rapid amplication of cDNA ends,RACE)的方法分离克隆了建鲤(Cyprinus carpiovar.Jian) PRL基因全长cDNA,得到1 028 bp的全长cDNA,包括633 bp的开放阅读框(ORF),51 bp 的5′末端非编码区(UTR)以及344 bp的3′末端非编码区(UTR)。对该基因序列和推测的氨基酸序列进行同源性比对和系统分析显示:建鲤与其它硬骨鱼类该基因的氨基酸序列相似度为55.02%~94.76%,与草鱼(Ctenopharyngodon idella)相似度最高(94.76%),与齐口裂腹鱼(Schizothorax prenanti)、斑马鱼(Danio rerio)相似度稍低,与红鳍东方鲀(Takifugu rubripes)的相似度较低(55.02%),表明不同物种间PRL的氨基酸序列具有较高的保守性。用实时定量PCR (RT PCR)检测该基因在建鲤脑垂体、脑、肠、鳃、性腺、肝、脾、肾中PRL的相对表达量,其中脑垂体最高,其次是肝、鳃、脾、脑、肠、肾,在性腺中也有较低的表达量,表明脑垂体是建鲤PRL基因的主要表达场所,在性腺、肝、脾的表达说明其在鱼体内除了渗透调控可能还存在多种生理功能。

    Abstract:

    To reveal the gene sequences of prolactin and its expression in osmotic regulation tissue of Cyprinus carpio var.Jian, the full length cDNA encoding prolactin was cloned from Cyprinus carpio var.Jian using homology cloning and RACE PCR approach. The prolactin was 1 028 bp in length, including 51 bp 5’terminal UTR, 633 bp encoding region and 344 bp 3’terminal UTR. Phylogenetic analysis using MEGA 4 software showed that the putative prolactin amino acid sequence and other teleosts have the high similarity of 55.02%-94.76%. The prolactin amino acid sequence of Cyprinus carpio var.Jian and that of Ctenopharyngodon idella were close to each other in similarity of 94.76%. For other fishes, the similarity was 94.26%,90.00%,70.53%,70.53%,65.70%,63.90%,63.29%,62.93%,62.38%,62.32%,55.02% with Schizothorax prenanti, Danio rerio, Salmo salar, Oncorhynchus mykiss, Anguilla japonica, Oreochromis Niloticus, Epinephelus coioides, Acanthopagrus schlegelii, Paralichthys olivaceus, Sparus aurata, Takifugu rubripes, respectively. Real time quantitative PCR results indicated that prolactin genes were expressed in the pituitary, brain, liver, kidney, intestines, gonad, spleen and gills. The prolactin transcript was detected at a high level in pituitary, moderate in brain, gills, spleen, liver, kidney and intestines, and low in gonad. This suggests that the pituitary gland is the main expression site of prolactin in Cyprinus carpio var.Jian.The prolactin transcript was detected in the gonads, liver, and spleen suggests that there may exist many physiological functions about prolactin in fish.

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王 杰,李 冰,张成锋,朱 健.建鲤催乳素基因全长cDNA的克隆及序列分析和组织表达[J].上海海洋大学学报,2012,21(6):935-944.
WANG Jie, LI Bing, ZHANG Cheng-feng, ZHU Jian. Full-length cDNA cloning and tissue expression of PRL from Cyprinus carpio var. Jian[J]. Journal of Shanghai Ocean University,2012,21(6):935-944.

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  • 在线发布日期: 2012-11-22
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