赤点石斑鱼FAS基因的克隆及序列分析

摘要

Objective] To research the structure and expression characteristics of FAS (Fatty Acid Synthase) gene in Epinephelus akaara. [ Method] With the transcriptome sequence obtained in previous research as the database , PCR technology was used to obtain the cDNA sequence of FAS gene in E.akaara.[Result] This sequence had high similarity with that of Larimichthy scrocea (87%).The total length of FAS gene was 8 505 bp, which included 712 bp 3′UTR, 7 548 bp open reading frame (ORF), and 245 bp 5′UTR.A total of 2 515 amino acids were enco-ded.The protein molecular weight was 274.03 ku, isoelectric point was 5.98, and it belonged to the hydrophilic protein.Results of Real-time PCR showed that FAS gene had the highest expression in heart, followed with liver.Evolution of FAS gene was in accord with that of species. [Conclusion] This research lays theoretical basis for the fat metabolism of E.akaara, and provides basic data for the cultivation of E.akaara.%[目的]研究赤点石斑鱼（ Epinephelus akaara） FAS基因的结构与表达特性。[方法]以前期获得的转录组序列作为数据库，应用PCR技术，获得赤点石斑鱼FAS 基因的cDNA全序列。[结果]该序列与大黄鱼（ Larimichthy scrocea） FAS基因的相似度最高，为87％。该序列全长8505 bp，包括712 bp的3′UTR区、7548 bp的开放阅读框（ ORF）以及245 bp的5′UTR区，可编码2515个氨基酸。经预测，该蛋白分子量为274．03 ku，等电点为5．98，其属于亲水性蛋白。通过Real-time PCR方法获得FAS在赤点石斑鱼各个组织中的表达，结果显示，其在心脏中表达量最高，其次是在肝脏中。 FAS基因的进化与物种进化一致。[结论]该研究为进一步研究赤点石斑鱼脂肪代谢奠定理论基础，也为赤点石斑鱼在养殖方面的工作提供基础资料。