| 李超琼，王雪芹，刘红占，等.凤丹牡丹PoFAD7基因的克隆及表达分析[J].中国油脂,2019,44(9):.[LI Chaoqiong, WANG Xueqin, LIU Hongzhan，etc.Cloning and expression analysis of PoFAD7 from Paeonia ostii[J].China Oils and Fats,2019,44(9):.] |
| 凤丹牡丹PoFAD7基因的克隆及表达分析 |
| Cloning and expression analysis of PoFAD7 from Paeonia ostii |
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| DOI： |
| 中文关键词: 凤丹牡丹 脂肪酸脱氢酶7 基因克隆 表达分析 |
| 英文关键词:Paeonia ostii fatty acid desaturase 7 gene clon expression analysis |
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| 中文摘要: |
| ω-3脂肪酸脱氢酶能将亚油酸催化合成ɑ-亚麻酸，是植物脂肪酸生物合成的关键酶。以凤丹牡丹（Paeonia ostii）种子为试验材料，采用RACE和RT-PCR方法克隆获得凤丹牡丹脂肪酸脱氢酶7基因cDNA片段，命名为PoFAD7（GenBank登录号：MK205360）；生物信息学分析表明其包含完整的开放阅读框，推测编码的蛋白质含有450个氨基酸，具有2个跨膜结构域。蛋白质氨基酸序列比对分析表明，PoFAD7含有3个保守的组氨酸基序，而系统进化树分析结果显示凤丹牡丹与同属芍药属的芍药亲缘关系较近。对不同发育时期种子中PoFAD7基因的表达分析结果表明，该基因在凤丹牡丹授粉100 d的种子中相对表达量最高，结合凤丹牡丹种子中亚油酸和α-亚麻酸的累积规律，推测PoFAD7在催化亚油酸合成α-亚麻酸的反应中发挥一定的功能，但可能并不是α-亚麻酸合成的主效基因。 |
| 英文摘要: |
| ω-3 Fatty acid desaturase is the key enzyme in the lipid biosynthesis, which has been documented to be responsible for producing α-linolenic acid from linoleic acid in plants. The fatty acid desaturase 7 gene of Paeonia ostii, named PoFAD7 (GenBank accession number： MK205360), was cloned using RACE and RT-PCR method. Bioinformatics analysis results showed that the PoFAD7 gene contained the complete cDNA open reading frame and encoded a protein with 450 amino acids and two transmembrane domains. Mutiple sequences alignment analysis indicated that PoFAD7 contained three histidine motifs, and phylogenetic tree analysis showed that Paeonia ostii had the closest evolutionary relationship with Paeonia lactiflora. Moreover, the expression analysis of PoFAD7 in seeds of different developmental stages showed that the highest expression level appeared at 100 d after pollination. Combining the results of the accumulation pattern of linoleic acid and α-linolenic acid, we speculated that PoFAD7 might play a role in the synthesis of α-linolenic acid from linoleic acid, but it was not the key gene in the synthesis of α-linolenic acid. |
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