摘要
CCAAT增强子结合蛋白α(C/EBPα)是脂肪细胞分化中起重要调控作用的转录因子。本研究利用逆转录PCR和RACE技术获得了体质量200~250 g鲤Cyprinus carpio C/EBPα基因全长cDNA序列为1463 bp,开放阅读框861 bp,编码286个氨基酸。C/EBPα蛋白质相对分子量32.5 KD,理论等电点(PI)为7.02。氨基酸同源性分析结果显示,鲤C/EBPα基因与斑马鱼Danio rerio同源性最高为93.36%,与小鼠同源性仅为52.47%。系统进化树结果显示,鲤C/EBPα氨基酸序列与鲫Carassius auratus聚为一支,同源性最高。实时荧光定量PCR检测表明,C/EBPα基因在成体鲤腹腔脂肪组织中表达量最高,肠组织次之,腹部肌肉中表达量最低;胚胎发育期,C/EBPα基因在心跳期表达量最高,七体节期次之,八细胞期最低。鲤C/EBPα基因序列的获得及表达特征研究为进一步深入研究鲤脂肪细胞分化及脂肪发育分子调控机制奠定了基础。
CCAAT/enhancer-binding protein(C/EBPα)gene as an important candidate gene for fat deposition plays a key role in the process of adipocyte differentiation.The full-length cDNA sequence of C/EBPαgene was cloned in common carp(Cyprinus carpio)by reverse transcription PCR and rapid-amplification of cDNA ends(RACE)technology,and the expression pattern of C/EBPαgene was investigated in different tissues of common carp during in different developmental stages.Full length of theC/EBPαcDNA was found to be 1,463 bp,with 5'untranslated region of 234 bp,3'untranslated region of 386 bp,and open reading frame(ORF)of 861 bp encoding 286 amino acids.The C/EBPαprotein had relative molecular weight of 32.5 KD,and theoretical PI value of 7.02,with fat coefficient of 63.50.Multiple sequences alignment revealed that the amino acid sequences of C/EBPαhad 93.36%identity with zebrafish Danio rerio,63.64%identity with Atlantic salmon Salmo salar,and 52.47%identity with mice Mus musculus.The phylogenetic tree revealed that C/EBPαgene clustered closely with herbivorous and omnivorous freshwater fish,most similar to crucian carp Carassius auratus.The real-time quantitative PCR showed that C/EBPαgene was expressed maximally in abdominal adipose tissue,followed by intestinal tissue in adult common carp and the minimal in muscular tissue.There was the maximal C/EBPαgene expression level in the heart beating period,followed by the seven myomere stage,and the minimal in the 8-cell stage,during the development of embryo.These findings will assist in further understanding of the biosynthesis of C/EBPαin common carp,and enable the methods to be developed for enhancing its production.
作者
孙志鹏
吕伟华
党红阳
曹顶臣
匡友谊
鲁翠云
郑先虎
SUN Zhipeng;Lü Weihua;DANG Hongyang;CAO Dingchen;KUANG Youyi;LU Cuiyun;ZHENG Xianhu(National Local Joint Engineering Laboratory for Freshwater Fish Breeding,Heilongjiang River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Harbin 150070,China;Key Laboratory of Freshwater Fishery Germplasm Resource,Ministry of Agriculture and Rural Affairs,Shanghai Ocean University,Shanghai 201306,China)
出处
《水产学杂志》
CAS
北大核心
2020年第6期22-27,共6页
Chinese Journal of Fisheries
基金
中央级公益性科研院所基本科研业务费专项(HSY201805M)
中国水产种质资源共享服务平台项目(2019DKA30470).
