摘要
采用RACE-PCR 克隆卵形鲳鲹(Trachinotus ovatus)过氧化物酶体增殖物激活受体(peroxisome proliferator activated receptors-α,PPAR α)基因的cDNA 序列全长,并应用生物信息学方法分析其编码蛋白质的理化性质和结构特征.结果表明:卵形鲳鲹PPAR α 基因(GenBank 登录号KP893147) cDNA 全长1 930 bp,开放阅读框(ORF)为1 425 bp,共编码474 个氨基酸,其编码的蛋白质为不稳定蛋白,无信号肽和跨膜结构,二级结构由α 螺旋、β转角、伸展片段和无规则卷曲组成,且α 螺旋占较大比例;预测显示,该蛋白有PPARs 基因家族典型的DNA 结合区(DBD)和配体结合区(LBD);序列对比表明,卵形鲳鲹PPAR α 基因与尼罗罗非鱼(Oreochromis niloticus)、花鲈(Lateolabrax japonicas)、大黄鱼(Larimichthys crocea)、金头鲷(Sparus aurata)、军曹鱼(Rachycentroncanadum)等有较高的同源性(81% - 89%);蛋白系统进化树分析显示,在人(Homo sapiens)、鼠(Mus musculus)、鸭(Gallus gallus)、花鲈、大黄鱼、军曹鱼等动物中,卵形鲳鲹的PPAR α 蛋白与军曹鱼的进化关系最为密切(94%),与人(68%)、鼠(68%)、鸭(67%)等的同源性较低.荧光定量分析显示,卵形鲳鲹PPAR α mRNA 在脑、肾脏、肠、脾脏等组织表达水平较高,其次是皮肤、肌肉,在心脏、肝脏中表达量较低.
The full-length c DNA of PPAR α in Trachinotus ovatus was cloned by RACE(Rapid Amplification of c DNA Ends), and the protein structures, physical, chemical properties and their functional domain structures which encoded by PPAR α gene were analyzed by bioinformatics method. The results showed that c DNA sequences of PPAR α in T. ovatus were 1 930 bp, and the open reading frame was 1 425 bp, encoding 474 amino acids(Gen Bank accession No. KP893147). The protein was unstable protein without signal peptide and trans-membrane. The secondary structure of PPAR α contains alpha helixes, beta turns, and extended strands, random coils. At the same time, alpha helixes were 45.15%. There were some typical structures in PPAR α, including a DNA-binding domain with zinc-finger structures, and a ligand binding domain which can integrate ligands and activate PPAR α. That the sequences of T. ovatus PPAR α were highly homologous with those cloned from Japanese sea perch, cobia, large yellow croaker and gilthead sea-bream(81%- 89%), phylogenetic tree analysis of PPAR α protein showed that the nearest relationship existed between T. ovatus and cobia among all the species mentioned above, and the lowest homology was human. Using quantitative real-time RT-PCR, we observed that PPAR α m RNA was expressed predominantly in brain-lipid, spleen, intestine and head-kidney tissues of T. ovatus.
出处
《广东海洋大学学报》
CAS
2015年第4期1-9,共9页
Journal of Guangdong Ocean University
基金
国家海洋公益性行业科研专项(201205028)
广东省海洋经济创新发展区域示范专项(GD2012-A01-007
GD2012-A02-003)
广东省教育厅创新计划专项(2012KJCX0063)
广西科技厅科技计划(桂科攻1222013-2)