黄鳝(Monopterus albus)肉碱棕榈酰转移酶基因克隆及组织分布分析  

Cloning and Tissue Distribution Analysis of Carnitine Palmyltransferase Gene in Swamp Eel(Monopterus albus)

作　　者：晁碧辉 陈喜 黄圣明 周秋白[1,2,3] 熊六凤 张亚洲[1,2,3] CHAO Bihui;CHEN Xi;HUANG Shengming;ZHOU Qiubai;XIONG Liufeng;ZHANG Yazhou(College of Animal Science and Technology,Jiangxi Agricultural University,Nanchang,330045;Nanchang Key Laboratory of Featured Hydrobios Nutritional Physiology and Healthy Breeding,Nanchang,330045;Research Institute of Special Fisheries,Jiangxi Agricultural University,Nanchang,330045)

机构地区：[1]江西农业大学动物科学技术学院,南昌330045 [2]南昌市特色水生生物营养生理与健康养殖重点实验室,南昌330045 [3]江西农业大学特种水产研究所,南昌330045

出　　处：《基因组学与应用生物学》2024年第3期453-464,共12页Genomics and Applied Biology

基　　金：江西省教育厅科技计划项目(GJJ200449);国家自然科学基金项目(32160869);国家特色淡水鱼产业技术体系(CARS-46);江西省大宗淡水鱼产业技术体系(JXARS-04)共同资助。

摘　　要：肉碱棕榈酰转移酶(carnitine palmitoyltransferas,CPT)是调节脊椎动物长链脂肪酸β-氧化的关键因子,包括肉碱棕榈酰转移酶1(carnitine palmitoyltransferas 1,CPT1)和肉碱棕榈酰转移酶2(carnitine palmitoyltransferas 2,CPT2),在能量代谢中起着重要调控作用。到目前为止,许多研究已经提供了在硬骨鱼中存在多种CPT基因的证据,但目前在黄鳝(Monopterus albus)中关于CPT基因的研究还未见报道。本研究以黄鳝肝脏cDNA为模板,采用巢式PCR的方法和TA克隆技术获得CPT1A、CPT1B和CPT2等3个基因的开放阅读框(open reading frame,ORF)序列,利用生物信息软件分析三者结构特征,并通过实时定量PCR检测其在不同组织间的表达情况。结果显示,CPT1A、CPT1B和CPT2基因的ORF序列长度分别为2367、2361和2004 bp,分别编码788、786和667个氨基酸;预测蛋白质成熟肽的分子量分别为89.83、89.70和74.55 kDa;3种蛋白的二级结构主要由α-螺旋构成。同源性分析发现,黄鳝CPT1A、CPT1B和CPT2蛋白与其他物种CPT蛋白间同源性较高,均大于60%;系统进化树显示黄鳝CPT1A、CPT1B和CPT2蛋白均与硬骨鱼棘鳍总目的亲缘关系较近。组织分布分析表明,黄鳝CPT1A和CPT1B在心脏中的表达量最高,CPT2在胃中表达量最高。通过对黄鳝CPT1A、CPT1B和CPT2基因进行表达分布研究,可推断它们可能在黄鳝营养代谢和性腺发育方面发挥作用。Carnitine palmitoyltransferase(CPT)is a key factor regulating theβ-oxidation of long-chain fatty acids in vertebrates,including carnitine palmitoyltransferase 1(CPT1)and carnitine palmitoyltransferase 2(CPT2),which plays an important role in energy metabolism.So far,many studies have provided evidence of a variety of CPT genes in bony fish,but the study of CPT gene in swamp eel(Monopterus albus)has not been reported.In this study,using the liver cDNA of swamp eel as a template,the open reading frame(ORF)sequences of CPT1A,CPT1B and CPT2 genes were obtained by nested PCR and TA cloning technique.The structural characteristics of the three genes were analyzed by bioinformatics software,and their expression in different tissues was detected by real time quantitative PCR.The results showed that the ORF sequences of CPT1A,CPT1B and CPT2 genes were 2367,2361,and 2004 bp,encoding 788,786,and 667 amino acids,respectively,and the molecular weights of predicted mature peptides were 89.83,89.70,and 74.55 kDa,respectively,and the secondary structures of the genes were mainly composed ofαhelix.Homology analysis showed that the CPT1A,CPT1B,and CPT2 protein of swamp eel had high homology with CPT proteins of the other species,all of which were more than 60%.The phylogenetic tree showed that the CPT1A,CPT1B,and CPT2 proteins of swamp eel were closely related to that of the spinous fin order of teleost.Tissue distribution analysis showed that the expression of CPT1A and CPT1B of swamp eel was the highest in heart,and the expression of CPT2 in stomach was the highest.By studying the expression and distribution of CPT1A,CPT1B,and CPT2 genes in swamp eel,we can infer that they may play a role in nutritional metabolism and gonadal development of swamp eel.

关 键 词：黄鳝 肉碱棕榈酰转移酶 克隆 组织分布 β-氧化 

分 类 号：S917.4[农业科学—水产科学]