三角褐指藻DGAT1基因生物信息学分析及表达调控研究  

Bioinformation analysis and expression regulation of DGAT1 gene in Phaeodactylum tricornutum

作　　者：王何瑜[1] 龚一富[2,3] 郑小恽 李申睿 WANG Heyu;GONG Yifu;ZHENG Xiaoyun;LI Shenrui(College of Food and Pharmaceutical Sciences,Ningbo University,Ningbo 315832,China;School of Marine Sciences,Ningbo University,Ningbo 315832,China;Key Laboratory of Marine Biotechnology of Zhejiang Province,Ningbo 315832,China)

机构地区：[1]宁波大学食品与药学学院,浙江宁波315832 [2]宁波大学海洋学院,浙江宁波315832 [3]浙江省海洋生物工程重点实验室,浙江宁波315832

出　　处：《宁波大学学报（理工版）》2021年第4期1-7,共7页Journal of Ningbo University：Natural Science and Engineering Edition

基　　金：浙江省科技厅重点科技创新团队项目(2012R10029-07,2010R50029);宁波市科技攻关项目(2014C91023,2013C10018);宁波市社发重大项目(2017C510002)。

摘　　要：本研究采用转录组测序获得了三角褐指藻DGAT1基因cDNA全长序列(GeneBank登录号:7200924),并对其进行生物信息学分析和表达调控研究.结果表明,三角褐指藻DGAT1基因cDNA序列全长为1 438 bp,开放阅读框(ORF)为1 098 bp,编码365氨基酸序列,含有脂肪酸蛋白特性(Ⅰ)和DAG结合位点(Ⅱ)等功能结构域.预测三角褐指藻DGAT1蛋白为亲水性蛋白,含有6个跨膜结构域和7个超强跨膜螺旋区,无信号肽.进化树分析表明,三角褐指藻与海链藻DGAT1蛋白同源性最高. RT-qPCR结果表明,光照强度和温度均显著促进三角褐指藻DGAT1基因的表达.随着光照强度和温度的增加,三角褐指藻DGAT1基因的表达量呈先升高后降低趋势,在光照强度为2 500 lx或温度为25℃时,三角褐指藻DGAT1基因的表达量达到最大,这与三角褐指藻总脂含量的变化趋势一致,同样揭示DGAT1蛋白与三角褐指藻总脂生物合成与积累密切相关.In the current study, the full-length cDNA sequence of DGAT1(GeneBank Accession Number: 7200924) in Phaeodactylum tricornutum was cloned by transcriptome sequencing. Bioinformatics analysis and expression difference were also studied. The results showed that the full length of the DGAT1 gene from P. tricornutum was 1 438 bp, had an open reading frame(ORF) of 1 098 bp with the fatty acid-binding protein signature(Ⅰ) and DAG binding site(Ⅱ) domains, encoding 365 amino acids. The protein prediction results showed that the DGAT1 protein from P. tricornutum was a hydrophilicity protein including 6 transmembrane structure domains and 7 super transmembrane helical regions, without signal peptide. Phylogenetic analysis results showed that DGAT1 protein from P. tricornutum had closed homologous relationship with the DGAT1 protein of Thalassiosira pseudonana. RT-qPCR results showed that light intensity and temperature could significantly promote the expression of DGAT1 gene of P. tricornutum. With the increase of light intensity and temperature, the expression quantity of DGAT1 gene from P. tricornutum showed an increased first and then decreased trend. The expression quantity of DGAT1 gene reached the maximum at light intensity of 2 500 lx or temperature of 25 ℃. The expression of DGAT1 gene was consistent with the total fat content. The results indicated that DGAT1 protein might be one of the key enzymes in the biosynthesis and accumulation of total lipid in P. tricornutum.

关 键 词：三角褐指藻 DGAT1基因 克隆 生物信息学 基因表达调控 

分 类 号：S917.3[农业科学—水产科学] Q943.2[生物学—植物学]