| 朱金鑫，孙金金，原晓龙，王娟，杨宇明，王毅.滇牡丹ω-3脂肪酸脱氢酶基因克隆与功能分析[J].中国油脂,2017,42(02):.[ZHU Jinxin,SUN Jinjin,YUAN Xiaolong,WANG Juan, YANG Yuming,WANG Yi.Cloning and functional analysis of omega-3 fatty acid desaturase gene from Paeonia delavayi[J].China Oils and Fats,2017,42(02):.] |
| 滇牡丹ω-3脂肪酸脱氢酶基因克隆与功能分析 |
| Cloning and functional analysis of omega-3 fatty acid desaturase gene from Paeonia delavayi |
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| DOI： |
| 中文关键词: 滇牡丹 ω-3脂肪酸脱氢酶 基因克隆 基因功能分析 |
| 英文关键词:Paeonia delavayi omega-3 fatty acid desaturase gene clone gene functional analysis |
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| 中文摘要: |
| ω-3脂肪酸脱氢酶是多不饱和脂肪酸亚麻酸生物合成途径的关键酶。依据转录组数据设计特异性引物，使用RT-PCR方法从滇牡丹克隆得到1个FAD3基因的cDNA全长序列，命名为PdFAD3(GeneBank登录号为KX289610)。生物信息学分析结果表明PdFAD3基因片段序列全长2 134 bp，包含完整的cDNA开放阅读框，编码含有450个氨基酸残基的蛋白质；Blast比对结果显示该蛋白质属于FAD蛋白质家族；系统进化树分析结果显示与芍药属芍药组植物芍药亲缘关系较近；该蛋白质属于跨膜蛋白质，亲水性较低；RT-PCR结果显示其在种子中的表达随着种子成熟出现双峰型的表达量变化，与目前所报道的其他物种的FAD3表达情况相似。为进一步研究其调控机理提供理论依据及获得含高不饱和脂肪酸转基因滇牡丹品种奠定理论基础。 |
| 英文摘要: |
| Omega-3 fatty acid desaturase is the key enzyme in the biosynthesis pathway of linolenic belonging to polyunsaturated fatty acids. A full-length cDNA sequences of a new FAD3 gene was successfully cloned from Paeonia delavayi by RT-PCR method with the specific primers designed based on transcriptome datas, named PdFAD3(GeneBank accession number: KX289610).Bioinformatics analysis results showed that full-length sequence of PdFAD3 gene was 2 134 bp, containing the complete cDNA open reading frame (ORF), encoded a protein with 450 amino acids. Blast comparison results showed that the protein belonged to the FAD family. Analysis results of phylogenetic tree showed that Paeonia delavayi had close genetic relationship with Paeonia lactiflora belonging to Paeonia Sect. Paeonia. The protein belonged to the transmembrane protein and was less hydrophilic. RT-PCR results showed that the expression in the seed as maturation occured bimodal change, similar to the FAD3 reported in other species. The PdFAD3 gene cloned from Paeonia delavayi provided a scientific basis for further study on its regulation mechanism and obtaining Paeonia delavayi transgenic variety with highly unsaturated fatty acids. |
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