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中华绒螯蟹脂肪酸延长酶(ELOVL)基因全长cDNA的克隆及其表达分析 [中文引用][英文引用]

Full-length cDNA cloning and expression analysis of the fatty acid elongase gene from Chinese mitten crab (Eriocheir sinensis)

作者(英文):YANG Zhigang  SHI Qiuyan  CHENG Yongxu  YAO Qinqin  QUE Youqing  YANG Qing  XU Lei  LIU Meimei  XU Zewen 
单位(英文):Shanghai Ocean University  Shanghai 201306  China 
关键词(英文):Eriocheir sinensis  elongases of very long chain fatty acids  gene clone  expression analysis 
分类号:S917
出版年·卷·期(页码):2016·23·第1期(53-63)
DOI: 10.3724/SP.J.1118.2016.15159
-----摘要:-------------------------------------------------------------------------------------------

根据脂肪酸延长酶保守区序列设计引物,采用RT-PCR和RACE技术首次克隆得到中华绒螯蟹(Eriocheir sinensis)脂肪酸延长酶(ELOVL)基因全长(GenBank登录号:KR005628)。分析ELOVL序列表明,该基因cDNA全长2089 bp,开放阅读框(ORF)长1065 bp(包含终止密码子),编码的354个氨基酸具有典型的延长酶特征:1个高度保守的氧化还原中心组氨酸簇HVIHH,多个保守区和多个跨膜区(KFTEFLDT、NTFVHIVMYVYY、TNFQMI)。经氨基酸同源性和系统发育树分析,中华绒螯蟹ELOVL预测氨基酸序列与顶切叶蚁(Acromyrmex echinatior)ELOVL氨基酸序列相似性最高,为59%;同时与家蝇(Musca domestica)聚为一支,进而与日本囊对虾(Marsupenaeus japonicus)等聚为一大支。通过实时荧光定量PCR技术研究ELOVL mRNA在中华绒螯蟹不同组织中的表达量,及投喂不同配合饲料后在可食部位组织中的表达情况。结果显示, ELOVL在各组织中均有表达,在肝胰腺和肠道中表达量最高,心脏中最低,其他组织中少量表达。养殖98 d后分析表明,卵巢和肝胰腺组织中ELOVL mRNA在SO饲料组中表达量最高,并且表达水平随着饲料中鱼油(富含n-3 PUFA)比例减少、豆油(缺少n-3 PUFA)比例增加而显著升高(P < 0.05);精巢组织中ELOVL mKNA表达量在SO饲料组中最高;肌肉组织中ELOVL mKNA表达水平较低,且各饲料组间表达量差异不显著(P > 0.05)。以上结果表明,中华绒螯蟹存在ELOVL基因,并且ELOVL的表达受饲料脂肪水平的影响,这为探究中华绒螯蟹自身合成HUFA途径及调节机制奠定了基础。

-----英文摘要:---------------------------------------------------------------------------------------

Highly unsaturated fatty acids (HUFA) play a particularly important role in normal growth, immune function, and reproduction. In this study, the full-length cDNA of fatty acid elongase (ELOVL) from Chinese mitten crab (Eriocheir sinensis) was cloned for the first time using reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends analyses. The ELOVL cDNA sequence (GenBank accession number: KR005628) was 2089 bp long, with a 350-bp 5'-untranslated region (UTR), a 674-bp 3'-UTR, and an open reading frame of 1065 bp, including stop codons that specified a protein of 354 amino acids. The presumed protein had a typical ELOVL structure with a diagnostic histidine box HVIHH motif, several conserved regions, and three putative transmembrane-spanning domains (KFTEFLDT, NTFVHIVMYVYY, and TNFQMI). The encoded protein shared 59% amino acid sequence identity with Acromyrmex echinatior (GenBank number: XP_011054336.1) and was clustered closely with Musca domestica and Marsupenaeus japonicus in a phylogenetic tree. The ELOVL gene was expressed in the hepatopancreas, intestine, muscle, stomach, heart, and gill. Higher levels were detected in the hepatopancreas and intestine (P < 0.05) and lower levels were observed in stomach, gill, cranial ganglia, muscle, thoracic ganglia, and the eyes, with trace levels expressed in the heart. We designed three different lipid-containing feeds and analyzed ELOVL mRNA expression in muscle, hepatopancreas, and gonads. The results showed the highest expression in the ovary after feeding for 98 days (P < 0.05). ELOVL transcripts increased in the ovary and hepatopancreas with increased inclusion of SO in the diet, but no expression differences were detected in muscle or testis. These results lay the foundation for further research into the mechanisms regulating E. sinensis HUFA biosynthesis.

-----参考文献:---------------------------------------------------------------------------------------

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若需在您的论文中引用此文,请按以下格式著录参考文献:
中文著录格式: 杨志刚,,施秋燕,,成永旭,,姚琴琴,,阙有清,,杨青,,徐蕾,,柳梅梅,,徐泽文.中华绒螯蟹脂肪酸延长酶(ELOVL)基因全长cDNA的克隆及其表达分析.中国水产科学.2016;23(1):53-63.
英文著录格式: YANG,Zhigang,,SHI,Qiuyan,,CHENG,Yongxu,,YAO,Qinqin,,QUE,Youqing,,YANG,Qing,,XU,Lei,,LIU,Meimei,,XU,Zewen.Full-length cDNA cloning and expression analysis of the fatty acid elongase gene from Chinese mitten crab (Eriocheir sinensis).No Title Settings.2016;23(1):53-63.

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