分子营养

鲈鱼视黄酸受体α和视黄酸受体γ cDNA克隆和基因表达分析

  • 钱云霞 ,
  • 韩柳 ,
  • 童彩环 ,
  • 郑伟贤
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  • 宁波大学生命科学与生物工程学院, 应用海洋生物技术教育部重点实验室, 宁波 315211

收稿日期: 2011-11-16

  网络出版日期: 2012-03-30

基金资助

国家自然科学基金资助项目(30671608)

Retinoic Acid Receptor α and Retinoic Acid Receptor γ in Lateolabrax japonicas: cDNA Cloning and Gene Expression Analysis

  • QIAN Yunxia ,
  • HAN Liu ,
  • TONG Caihuan ,
  • ZHENG Weixian
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  • Key Laboratory of Applied Marine Biotechnology of Ministry of Education, Faculty of Life Science and Biotechnology, Ningbo University, Ningbo 315211, China

Received date: 2011-11-16

  Online published: 2012-03-30

摘要

本文旨在研究鲈鱼视黄酸受体α(RARα)和视黄酸受体γ(RARγ)的结构及其基因的组织表达特点。采用反转录PCR和cDNA末端快速克隆(RACE)技术,从鲈鱼肝脏中克隆得到RARα和RARγ全长cDNA序列。检测鲈鱼肝脏、肌肉、心脏、眼、肠、肾脏、脂肪、脾脏、鳃和大脑10个组织中RARαRARγ基因的表达情况。结果表明:1)鲈鱼RARα cDNA全长2 094 bp,5'端和3'端的非翻译区分别为124和608 bp,开放阅读框为1 362 bp,推测编码453个氨基酸,分子质量为50.64 ku,理论等电点为8.20。2)RARγ cDNA全长1 671 bp,其中包括36 bp的5'端非翻译区、129 bp的 3'端的非翻译区和1 506 bp的开放阅读框,共编码501个氨基酸,分子质量为56.20 ku,理论等电点为4.96。3)鲈鱼RARα和RARγ都具有典型的核受体结构,两者氨基酸序列同源性高达63.1%。鲈鱼的RARα和RARγ与红鳍东方鲀有较高的同源性,分别为96.9%和95.2%。4)RARαRARγ基因在所有检测组织中均有表达,其中RARα基因在心脏和大脑中表达较少,在眼、鳃和肠中表达较高,而RARγ基因仅在鳃和脾脏中有较高表达。总之,本试验成功克隆了鲈鱼RARα和RARγ的全长cDNA;鲈鱼RARα和RARγ有着典型的核受体结构,在各组织中广泛分布。

本文引用格式

钱云霞 , 韩柳 , 童彩环 , 郑伟贤 . 鲈鱼视黄酸受体α和视黄酸受体γ cDNA克隆和基因表达分析[J]. 动物营养学报, 2012 , 24(4) : 712 -721 . DOI: 10.3969/j.issn.1006-267x.2012.04.016

Abstract

This study was conducted to investigate structures of retinoic acid receptor α (RARα) and retinoic acid receptor γ (RARγ) and gene expression profiles in tissues. Reverse transcription PCR (RT-PCR) and rapid amplification of cDNA ends (RACE) methods were applied for the cloning of full-length cDNA coding for RARα and RARγ. Gene expressions of RARα and RARγ in various tissues, such as liver, muscles, heart, eyes, intestine, kidney, fat, spleen, gill and brain, were determined. The results showed as follows: 1) full-length cDNA coding for RARα was 2 094 bp, which included a 124 bp of 5'- untranslated region (UTR), a 608 bp 3'-UTR and a 1 362 bp open reading frame (ORF), the predicted RARα consisted of 453 amino acids with a molecular weight of 56.20 ku, and the theoretical isoelectric point was 8.20. 2) Full-length cDNA coding for RARγ was 1 671 bp consisting of a 36 bp 5'-UTR, a 129 bp 3'-UTR and a 1 506 bp ORF, 501 amino acids with a theoretical isoelectric point of 4.96 and a molecular weight of 56.20 ku was coded. 3) Both RARα and RARγ had typical nuclear receptor structure, and they shared 63.1% identity in their amino acid sequences. Lateolabrax japonicus RARα and RARγ shared high identity (96.9% and 95.2%, respectively) with those of Takifugu rubripes. 4) Lateolabrax japonicus RARα and RARγ gene expressions were detected in all the tested tissues, the higher gene expressions of RARα were in eyes, gill and intestine, the lower values were in heart and brain, and the higher gene expressions of RARγ were in gill and spleen. In conclusion, full-length cDNA of RARα and RARγ in Lateolabrax japonicas are successfully cloned in this study, typical nuclear receptor structure is contained both in RARα and RARγ, and they show ubiquitous tissue distribution characteristics.

参考文献

[1] CLAGETT-DAME M,DELUCA H F.The role of vitamin A inmammalian reproduction and embryonic development[J].Annual Review of Nutrition,2002,22:347-381.

[2] MOREN M,OPSTAD I,HAMRE K,et al.An optimum level of vitamin A supplements for Atlantic halibut (Hippoglossus hippoglossus L.) juveniles[J].Aquaculture,2004,235:587-599.

[3] HEMRE G I,DENG D F,WILSON R P,et al.Vitamin A metabolism and early biological responses in juvenile sunshine bass (Morone chrysops×M.saxatilis) fed graded levels of vitamin A[J].Aquaculture,2004,235:645-658.

[4] YANG Q H,ZHOU X Q,JIANG J,et al.Effect of dietary vitamin A deficiency on growth performance,feed utilization and immune responses of juvenile Jian carp (Cyprinus carpio var.Jian)[J].Aquaculture Research,2008,39:902-906.

[5] DUESTER G.Retinoic acid synthesis and signaling during early organogenesis[J].Cell,2008,134(6):921-931.  

[6] LIU R Z,SHARMA M K,SUN Q,et al.Retention of the duplicated cellular retinoic acid-binding protein 1 genes (crabp1a and crabp1b) in the zebrafish genome by subfunctionalization of tissue-specific expression[J].Federation of European Biochemical Societies,2005,272(14):3561-3571.

[7] ROCHETTE-EGLY C,GERMAIN P.Dynamic and combinatorial control of gene expression by nuclear retinoic acid receptors (RARs)[J].Nuclear Receptor Signaling,2009,7:e005.

[8] DELACROIX L,MOUTIER E,ALTOBELLI G,et al.Cell-specific interaction of retinoic acid receptors with target genes in mouse embryonic fibroblasts and embryonic stem cells[J].Molecular Cell Biology,2010,30:231-244.

[9] BASTIEN J,ROCHETTE-EGLY C.Nuclear retinoid receptors and the transcription of retinoid-target genes[J].Gene,2004,328:1-16.

[10] GERMAIN P,STAELS B,DACQUET C,et al.Overview of no menclature of nuclear receptors[J].Pharmacological Reviews,2006,58:685-704.

[11] KOIDE T,DOWNES M,CHANDRARATNA R A,et al.Active repression of RAR signaling is required for head formation[J].Genes & Development,2001,15:2111-2121.

[12] NAGASHIMA M,SAKURAI H,MAWATARI K,et al.Involvement of retinoic acid signaling in goldfish optic nerve regeneration[J].Neurochemistry International,2009,54:229-236.

[13] LINVILLE A,RADTKE K,WAXMAN J S,et al.Combinatorial roles for zebrafish retinoic acid receptors in the hindbrain,limbs and pharyngeal arches[J].Developmental Biology,2009,325:60-70.

[14] WENTWORTH J M,SCHOENFELD V,MEEK S,et al.Isolation and characterisation of the retinoic acid receptor-alpha gene in the Japanese pufferfish,F.rubripes[J].Gene,1999,236(2):315-323.  

[15] HALE A,TALLAFUSS A,YAN Y L,et al.Characterization of the retinoic acid receptor genes raraa,rarab and rarg during zebrafish development[J].Gene Expression Patterns,2006,6(5):546-555.  

[16] PERLMANN T,RANGARAJAN P N,UMESONO K,et al. Determinants for selective RAR and TR recognition of direct repeat HREs[J].Genes & Development,1993,7:1411-1422.

[17] CHAMBON P.A decade of molecular biology of retinoic acid receptors[J].Federation of American Societies for Experimental Biology,1996,10:940-954.

[18] RENAUD J P,ROCHEL N,RUFF M,et al.Crystal structure of the RAR-γ ligand-binding domain bound to all-trans retinoic acid[J].Nature,1995,378:681-689.

[19] MENG Q Y,CHEN X N,ZHAO J,et al.Distribution of retinoic acid receptor-α immunoreactivity in the human hypothalamus[J].Neuroscience,2011,174:132-142.

[20] DAY R M,LEE Y H,PARK A M,et al.Retinoic acid inhibits airway smooth muscle cell migration[J].American Journal of Respiratory Cell and Molecular Biology,2006,34:695-703.

[21] DOLLE P,RUBERTE E,LEROY P,et al.Retinoic acid receptors and cellular retinoid binding proteins.Ⅰ.A systematic study of their differential pattern of transcription during mouse organogenesis[J].Development,1990,110:1133-1151.

[22] LUFKIN T,LOHNES D,MARK M,et al.High postnatal lethality and testis degeneration in retinoic acid receptor α mutant mice[J]. Proceedings of the National Academy of Sciences,1993,90:7225-7229.

[23] CHAPELLIER B,MARK M,MESSADDEQ N,et al.Physiological and retinoid-induced proliferations of epidermis basal keratinocytes are differently controlled[J].The EMBO Journal,2002,21:3402-3413.
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