紫苏二酰甘油酰基转移酶基因(PfDGAT1)鉴定及表达分析

Gene Identification and Expression Analysis of Diacylgycerol Acyltransferase Gene(PfDGAT1) from Perilla frntescens

二酰甘油酰基转移酶(DGAT)是TAG合成过程中的限速酶。以晋紫苏1号为材料,采用电子克隆技术获得DGAT1基因(PfDGAT1,AF298815.1),并对PfDGAT1基因序列进行生物信息学及表达特性分析。结果表明,PfDGAT1基因序列全长为1 964 bp,开放阅读框长度为1 605 bp,共编码534个氨基酸残基;多序列比对和系统进化树分析表明,紫苏Pf DGAT1蛋白与芝麻、油橄榄的DGAT1蛋白的亲缘关系较近,序列同源性分别为89%和83%;利用实时荧光定量PCR技术分析PfDGAT1基因在紫苏不同组织中的表达特性,结果显示,PfDGAT1基因在紫苏不同组织中均有表达,但在种子中表达量最高,且随种子发育表达量呈先升高后降低的变化趋势,在开花后20 d表达量达到最高。

Diacylgycerol acyltransferase(DGAT) is the key enzyme and rate-limiting enzyme in the synthesis process of TAG. In this study, the PfDGAT1 gene(PfDGAT1, AF298815.1) was obtained from Jinzisu 1 by electronic cloning, and the bioinformatics and expression characteristics of PfDGAT1 gene sequence were analyzed. The bioinformatics analysis of the PfDGAT1 gene sequence showed that PfDGAT1 gene full length sequence was 1 964 bp. It contained 1 605 bp open reading frame (ORF) which encoded 534 amino acid residues. Multiple sequence alignments and phylogenetic analysis revealed that the deduced amino acid sequences of PfDGAT1 gene had close genetic relationship with its homologs sequences from Sesamum indicum and Olea europaea, and the sequence similarity was up to 89% and 83%, respectively. The expression characteristics of PfDGAT1 gene in different tissues of Perilla frutescens were investigated by real-time quantitative PCR. The results showed that PfDGAT1 gene was expressed in different tissues of Perilla frutescens, and had the highest expression level in seeds. The expression level of PfDGAT1 increased first and then decreased with the advancement of seed development stage, and reached the maximum at 20 d after flowering.