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

作　　者：朱金鑫[1,2] 孙金金[1,2] 原晓龙[2,3] 王娟[2,3] 杨宇明[2,3] 王毅[2,3] 

机构地区：[1]西南林业大学林学院,昆明650224 [2]国家林业局重点开放性实验室云南珍稀濒特森林植物保护和繁育实验室云南省森林植物培育与开发利用重点实验室,昆明650204 [3]云南省林业科学院,昆明650204

出　　处：《中国油脂》2017年第2期102-106,共5页China Oils and Fats

基　　金：云南省林业科学院重点实验室自主创新项目(22CX2016-05);云南省科技计划项目(2015A005);云南省应用基础研究重点项目(2013FA054)

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c DNA全长序列,命名为Pd FAD3(Gene Bank登录号为KX289610)。生物信息学分析结果表明Pd FAD3基因片段序列全长2 134 bp,包含完整的c DNA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。Omega -3 fatty acid desaturase is the key enzyme in the biosynthesis pathway of linolenic be- longing to polyunsaturated fatty acids. A full - length eDNA sequences of a new FAD3 gene was success- fully cloned from Paeonia delavayi by RT - PCR method with the specific primers designed based on tran- seriptome datas, named PdFAD3 （GeneBank accession number： KX289610）. Bioinformaties analysis re- suits showed that full -length sequence of PdFAD3 gene was 2 134 bp, containing the complete eDNA 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 phylogenetie 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 otherspecies. 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.

关 键 词：滇牡丹 ω-3脂肪酸脱氢酶 基因克隆 基因功能分析 

分 类 号：S685.11[农业科学—观赏园艺]