| 日本鳗鲡脂肪酸去饱和酶和延长酶基因的克隆与分析 |
| Molecular Cloning and Analyzing of Fatty Acid Desaturase and Elongase Genes in Japanese Eel (Anguilla japonica) |
| 投稿时间：2008-12-03 |
| DOI： |
| 中文关键词:脂肪酸去饱和酶 脂肪酸延长酶 基因克隆 日本鳗鲡 |
| 英文关键词:Fatty acid desaturase Fatty acid elongase Gene cloning Japanese eel （AnguiUa japonica） |
| 基金项目: |
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| 中文摘要: |
| 利用RT—PCR和RACE方法克隆得到日本鳗鲡（Anguilla japonica）肝脏中控制高不饱和脂肪酸（highly unsaturated fatty acids，HUFA）合成的脂肪酸去饱和酶（fatty acid desaturase，FAD）和脂肪酸延长酶（fatty acid elongase，ELO）全长cDNA序列。结果表明，2456bp的FAD全长cDNA序列含长达1046bp的3’-UTR、75bp的5’-UTR和编码444个氨基酸的长1335bp的阅读框。编码的蛋白序列含有FAD全部的特征结构区，包括3个组氨酸簇、2个跨膜区和1个细胞色素h5结构域，与其他具有不同生活史鱼类的FAD氨基酸序列具有70．5％～77．5％的同源性；分析显示其在系统树中与溯河洄游鱼类的亲缘关系最近。克隆得到的ELO全长cDNA序列长1239bp，含有885bp的开放阅读框，编码294个氨基酸，该蛋白序列含有单一的氧化还原中心组氨酸簇、内质网停留信号和多个跨膜区等ELO特征结构，与其他鱼类ELO氨基酸具有87．1％～88．8％的同源性；其在系统树中与北非鲶鱼（Clarias gariepinus）和斑马鱼亲缘关系较近。日本鳗鲡FAD和ELOcDNA全序列的获得为今后进一步研究其体内高不饱和脂肪酸合成途径及阐明该途径在不同鱼类中的分子进化机理奠定基础。 |
| 英文摘要: |
| Full-length cDNAs of fatty acid desaturase （FAD） and fatty acid elongase （ELO）, which were involved in the biosynthesis of highly unsaturated fatty acids （ HUFA）, were isolated from an anadromous teleost fish Japanese eel （Anguilla japonica） by RT-PCR and RACE methods. The results revealed that the FAD cDNA was 2 456 bp, including a 3＇ - UTR as long as 1 046 bp, a 5＇ - UTR of 75 bp and a 1 335 bp ORF specifying a protein of 444 amino acids. The protein sequence contained all the characteristics of microsomal fatty acid desaturases, including three histidine boxes, two transmembrane regions, and one cytochrome b5 domain. The deduced amino acid sequence shared 70.5% -77.5% identity with other fish of different life history and it had a closer genetic relationship with anadromous migratory fish in the phylogenetic tree. The 1 239 bp cDNA of ELO included an 885 bp ORF encoding a 294 amino acid peptide that contained one single histidine box redox centre motif, one canonical ER retention signal and several transmembrane regions, in line with other fatty acid elongases. The deduced protein sequence had 87.1% - 88.8% identity with other fish and the phylogenetic tree showed that it had a closer genetic relationship with North African catfish （Clarias gariepinus） and zebrafish. These results will help study the relationship between their structure and function, and elucidate the HUFA biosynthesis pathway in Japanese eel. |
| 郁 颖,梁旭方,李观贵,何珊,谢骏,白俊杰.2009.日本鳗鲡脂肪酸去饱和酶和延长酶基因的克隆与分析[J].水生态学杂志,30(5):56-62. |
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