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斑点叉尾鮰中阿黑皮素原cDNA的分子克隆及多组织mRNA表达

Molecular cloning of proopiomelanocortin cDNA and multi-tissue mRNA expression in channel catfish.

作者信息

Karsi Attila, Waldbieser Geoffrey C, Small Brian C, Liu Zhanjiang, Wolters William R

机构信息

USDA-ARS Catfish Genetics Research Unit, Thad Cochran National Warmwater Aquaculture Center, P.O. Box 38, Stoneville, MS 38776, USA.

出版信息

Gen Comp Endocrinol. 2004 Jul;137(3):312-21. doi: 10.1016/j.ygcen.2004.03.012.

Abstract

Channel catfish (Ictalurus punctatus) proopiomelanocortin (POMC) cDNA was cloned to investigate its structure, evolution, and expression in different tissues. POMC is an important gene in the hypothalamus-pituitary-adrenal axis, the main mediator of the stress response. POMC gene was isolated from a pituitary cDNA library and nucleotide sequence was determined. POMC cDNA is composed of 1164 nucleotides with a 639 nucleotide open reading frame encoding a protein of 212 amino acids. Catfish POMC protein contains a signal peptide (SP, Met(1)-Ala(28)), N-terminal peptide (Gln(29)-Glu(101)), adrenocorticotropic hormone (ACTH, Ser(104)-Met(142)), alpha-melanocyte stimulating hormone (alpha-MSH, Ser(104)-Val(116)), corticotropin-like intermediate lobe peptide (CLIP, Arg(121)-Met(142)), beta-lipotropin (beta-LPH, Glu(145)-His(212)), gamma-lipotropin (gamma-LPH, Glu(145)- Ser(177)), beta-MSH (Asp(161)-Ser(177)), and beta-endorphin (beta-EP, Tyr(180)-His(212)). Catfish POMC protein does not contain a gamma-MSH region and most of the joining peptide and part of the gamma-LPH are deleted. Protein sequence alignment showed the highest similarity with the carp (Cyprinus carpio) POMC I (66.5%) and POMC II (67%), while the sea lamprey (Petromyzon marinus) POC (17.9%) and POM (18.8%) were the most divergent. The average similarity was 46.95% among the 44 POMC proteins from 36 different species analyzed. Compared to the POMC mRNA levels in the pituitary, the concentration of the POMC mRNA was 0.0594% in the anterior kidney and 0.0012-0.0045% in all the other tissues except in the skin where the lowest expression (0.0005%) was observed. Overall architecture of channel catfish POMC is highly similar to those from other teleosts.

摘要

克隆了斑点叉尾鮰(Ictalurus punctatus)的阿黑皮素原(POMC)cDNA,以研究其结构、进化以及在不同组织中的表达。POMC是下丘脑-垂体-肾上腺轴中的一个重要基因,是应激反应的主要调节因子。从垂体cDNA文库中分离出POMC基因并测定了其核苷酸序列。POMC cDNA由1164个核苷酸组成,有一个639个核苷酸的开放阅读框,编码一个212个氨基酸的蛋白质。斑点叉尾鮰POMC蛋白包含一个信号肽(SP,Met(1)-Ala(28))、N端肽(Gln(29)-Glu(101))、促肾上腺皮质激素(ACTH,Ser(104)-Met(142))、α-黑素细胞刺激素(α-MSH,Ser(104)-Val(116))、促肾上腺皮质激素样中叶肽(CLIP,Arg(121)-Met(142))、β-促脂素(β-LPH,Glu(145)-His(212))、γ-促脂素(γ-LPH,Glu(145)-Ser(177))、β-黑素细胞刺激素(β-MSH,Asp(161)-Ser(177))和β-内啡肽(β-EP,Tyr(180)-His(212))。斑点叉尾鮰POMC蛋白不包含γ-MSH区域,并且大部分连接肽和部分γ-LPH被删除。蛋白质序列比对显示,与鲤鱼(Cyprinus carpio)的POMC I(66.5%)和POMC II(67%)相似度最高,而海七鳃鳗(Petromyzon marinus)的POC(17.9%)和POM(18.8%)差异最大。在所分析的36个不同物种的44种POMC蛋白中,平均相似度为46.95%。与垂体中的POMC mRNA水平相比,前肾中POMC mRNA的浓度为0.0594%,除皮肤中观察到最低表达(0.0005%)外,所有其他组织中的浓度为0.0012 - 0.0045%。斑点叉尾鮰POMC的整体结构与其他硬骨鱼类的高度相似。

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