斜带石斑鱼肝脂酶和脂蛋白脂酶基因克隆与序列分析  

Cloning and sequence analysis of hepatic lipase (HL) and liportein lipase (LPL) gene in Orange-Spotted Grouper(Epinephelus coioides)

作　　者：胡永乐[1] 梁旭方[1] 李观贵[1] 王琳[1] 黄志东[1] 

机构地区：[1]暨南大学生物工程学系,广东广州510632

出　　处：《暨南大学学报（自然科学与医学版）》2010年第5期520-527,共8页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：国家科技部863项目(2007AA09Z437);国家自然科学基金项目(30670367);广东省科技计划项目(2007B020701002;2005B20301005);广东省自然科学基金项目(031886)

摘　　要：为从分子水平研究海水鱼类肝脂酶(hepatic lipase,HL)和脂蛋白脂酶(liportein lipase,LPL)基因在脂质代谢过程中的作用及其分子系统进化地位,采用RT-PCR及RACE法从斜带石斑鱼(Epinephelus coioides)肝脏克隆得到HL、LPL基因cDNA全序列.克隆获得斜带石斑鱼肝脏HL基因cDNA全长2 261 bp,其中5′非翻译区(5′-UTR)为186 bp,3′-UTR为590 bp,开放阅读框(ORF)为1 485 bp,编码494个氨基酸;LPL基因cDNA全长2 191 bp,其中5′-UTR为144 bp,3′-UTR为499 bp,ORF为1 548 bp,编码515个氨基酸.序列同源性分析表明,斜带石斑鱼HL、LPL基因与哺乳动物同源性均低于真骨鱼类罗非鱼、斑鳢和真鲷等,这与其亲缘远近关系相一致,表明HL、LPL在进化过程中都相对保守.构建系统发生树显示,斜带石斑鱼HL、LPL基因氨基酸序列与大口黑鲈、真鲷等共同占据进化树上独立的分枝,与较为原始的硬骨鱼类中华鲟在进化树上距离较远.本实验对斜带石斑鱼HL、LPL基因同源性、系统进化地位的研究,为进一步研究海水鱼类脂代谢调控机制,预防鱼类脂代谢疾病奠定了基础.To research the molecular mechanism of hepatic lipase （HL） and liportein lipase （LPL） genes in lipid metabolism as well as their molecular evolutional relationship, the full-length cDNA sequences encoding HL and LPL were obtained from the liver of orange-spotted grouper （ Epinephelus eo- ioides） by RT-PCR and RACE methods. The sequence analysis showed that the complete eDNA se- quence of HL of orange-spotted grouper was 2261bp in length, containing a coding region of 1485bp （encoding coding 494 amino acids）. And the complete cDNA sequence of LPL was 2191bp in length, containing a region of 1548bp （encoding 515 amino acids）. Sequence analysis reveals that, the identity of conserved amino acid sequence of HL and LPL revealed the similarity of mammals was both lower than teleosts later in the evolution, the identities of HL and LPL amino acid sequence among mammals and fishes were relatively conserved. Phylogenetic analyses revealed that, the deduced amino acid sequence of the orange-spotted grouper was more similar to largemouth bass and red sea bream than Chinese Sturgeon. This research provides reliable evidences about the identity of amino acid sequence of HL and LPL and is useful for investigating physiology mechanism in relation to lipid metabolism and help to prevent beforehand lipid metabolism diseases in marine fish.

关 键 词：斜带石斑鱼 肝脂酶 脂蛋白脂酶 基因克隆 序列分析 

分 类 号：Q786[生物学—分子生物学]