紫苏脂肪酸硫酯酶基因PfFatA生物信息学及其表达特性分析
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摘要
为研究脂肪酸硫酯酶A(FatA)在紫苏脂肪酸合成机制中发挥的作用,对紫苏FatA基因及其编码的蛋白质进行生物信息学分析。结果表明,PfFatA基因cDNA全长1 652 bp,开放阅读框1 119 bp,编码372个氨基酸;紫苏FatA基因编码的蛋白质为亲水性不稳定蛋白,无跨膜区域;含Acyl-ACP_TE保守结构域,属于Hotdog fold超蛋白家族;多序列比对结果发现,序列C端含有保守的三联肽AKL和SKV结构;系统进化树分析表明,紫苏与丹参亲缘关系较近。通过Real-time PCR分析得知,PfFatA在紫苏根、茎、叶、花和种子不同发育时期均有表达,其中,在种子发育初期表达量最高。该研究结果可为进一步阐明PfFatA基因的功能及作用机制奠定基础。
In order to detect the function of fatty acid thioesterase(FatA) in fatty acid synthesis mechansim of perilla,many bioinformatics analytical methods were used to analyze the protein encoded byFatA gene. The result showed that full length of cDNA for PfFatA was 1 652 bp,open reading frame(ORF) was 1 119 bp,coding 372 amino acids.FatA was an unstable hydrophilic protein,without transmembrane region.FatA protein contained a conserved domain of Acyl-ACP_TE,belonging to the Hotdog fold super protein family. Multiple sequence alignment indicated thatFatA had highly conserved triple peptide of AKL,SKV in the C-terminal. Phylogenetic tree analysis showed that Perilla frutescens had the closest genetic relationship with Salvia miltiorrhiza. The result of Real-time PCR showed PfFatA was expressed in roots,stems,leaves,flowers and seeds from different development period,especially in seeds formed in 10 days after flowering. These results of the current study provided a foundation for further illustrating the function and mechanism of PfFatA gene.
In order to detect the function of fatty acid thioesterase(FatA) in fatty acid synthesis mechansim of perilla,many bioinformatics analytical methods were used to analyze the protein encoded byFatA gene. The result showed that full length of cDNA for PfFatA was 1 652 bp,open reading frame(ORF) was 1 119 bp,coding 372 amino acids.FatA was an unstable hydrophilic protein,without transmembrane region.FatA protein contained a conserved domain of Acyl-ACP_TE,belonging to the Hotdog fold super protein family. Multiple sequence alignment indicated thatFatA had highly conserved triple peptide of AKL,SKV in the C-terminal. Phylogenetic tree analysis showed that Perilla frutescens had the closest genetic relationship with Salvia miltiorrhiza. The result of Real-time PCR showed PfFatA was expressed in roots,stems,leaves,flowers and seeds from different development period,especially in seeds formed in 10 days after flowering. These results of the current study provided a foundation for further illustrating the function and mechanism of PfFatA gene.
引文
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[12]邱建丁,梁汝萍,谭学才,等.膜蛋白跨膜区段的预测分析[J].高等学校化学学报,2004(5):831-836.
[13]谭晓风,王建勇,龙洪旭,等.油茶脂酰辅酶A硫酯酶基因的克隆与表达分析[J].植物生理学报,2014,50(12):1845-1856.
[14]Buch C,Hunt M C,Alexson S E H,et al.Localization of peroxisomal matrix protein by photobleaching[J].Biochem Bioph Res Co,2009,388(2):355-359.
[15]王丽梅,叶诚,吴晨,等.紫苏油对衰老模型大鼠的抗衰老作用研究[J].食品科技,2013(1):280-284.
[16]周奕华,陈正华.植物种子中脂肪酸代谢途径的遗传调控与基因工程[J].植物学通报,1998,15(5):16-23.
[17]李会珍,孙子文,李晓君,等.紫苏种子性状与主要营养成分相关性分析[J].中国粮油学报,2013,28(10):55-59.
[18]Zheng H,Rowland O,Kunst L.Disruptions of the Arabidopsis enoyl-co A reductase gene reveal an essential role for very-long-chain fatty acid synthesis in cell expansion during plant morphogenesis[J].Plant Cell,2005,17(5):1467-1481.
[19]曾艳玲,谭晓风,张党权,等.油茶脂肪酸代谢途径中关键酶基因调控油脂合成的规律研究[J].中国粮油学报,2014,29(2):26-29,35.
[20]Chen G,Peng Z Y,Shan L,et al.Functional expression analysis of an acy-ACP thioesterase Fat B1 from Arachis hypogaea L.seed in Escherichia coli[J].Journal of food,Agriculture&Environment,2012,10(3):332-336.
[2]Jones A,Davies H M,Voelker T A.Palmitoyl-acyl carrier protein(ACP)thioesterase and the evolutidnary-origin of plant acyl-ACP thioesterases[J].Plant Cell,1995,7(3):359-371.
[3]Sergei Mekhedov,Oskar Martinez de Ilarduya,John Ohlrogge.Toward a functional catalog of the plant genome.A survey of genes for lipid biosynthesis[J].Plant Physiol,2000,122(2):389-402.
[4]Beisson F,Koo A J K,Ruuska S,et al.Arabidopsis genes involved in acyl lipid metabolism.A 2003 census of the candidates,a study of the distribution of expressed sequence tags in organs,and a web-based database[J].Plant Physiol,2003,132(2):681-697.
[5]Wang Q,Huang W,Jiang Q,et al.Lower levels of expression of FATA2 gene promote longer siliques with modified seed oil content in Arabidopsis thaliana[J].Plant Mol Biol Rep,2013,31(6):1368-1375.
[6]张晓彬,姜文鑫,张琳,等.紫苏的研究进展[J].食品研究与开发,2015,(7):140-143.
[7]Napier J A,Haslam R P,Beaudoin F,et al.Understanding and manipulating plant lipid composition:metabolic engineering leads the way[J].Curr Opin Plant Biol,2014,19(1):68-75.
[8]李铁柱,杜红岩,刘攀峰.杜仲α-亚麻酸生物合成相关基因的差异表达[J].经济林研究,2013,31(4):30.
[9]刘志勇,王孝宜,高建昌,等.番茄S-腺苷蛋氨酸脱羧酶基因SLSAMDC1的克隆与序列分析[J].园艺学报,2008,35(8):1137-1146.
[10]王计平,张玲慧,赵静,等.紫苏种子脂肪酸代谢及关键酶基因调控油脂合成规律的研究[J].中国粮油学报,2016,31(3):91-95.
[11]Livak K J,Schmittgen T D.Analysis of relative gene expression data using real-time quantitative PCR and the2-ΔΔCTmethod[J].Methods,2001,25(4):402-408.
[12]邱建丁,梁汝萍,谭学才,等.膜蛋白跨膜区段的预测分析[J].高等学校化学学报,2004(5):831-836.
[13]谭晓风,王建勇,龙洪旭,等.油茶脂酰辅酶A硫酯酶基因的克隆与表达分析[J].植物生理学报,2014,50(12):1845-1856.
[14]Buch C,Hunt M C,Alexson S E H,et al.Localization of peroxisomal matrix protein by photobleaching[J].Biochem Bioph Res Co,2009,388(2):355-359.
[15]王丽梅,叶诚,吴晨,等.紫苏油对衰老模型大鼠的抗衰老作用研究[J].食品科技,2013(1):280-284.
[16]周奕华,陈正华.植物种子中脂肪酸代谢途径的遗传调控与基因工程[J].植物学通报,1998,15(5):16-23.
[17]李会珍,孙子文,李晓君,等.紫苏种子性状与主要营养成分相关性分析[J].中国粮油学报,2013,28(10):55-59.
[18]Zheng H,Rowland O,Kunst L.Disruptions of the Arabidopsis enoyl-co A reductase gene reveal an essential role for very-long-chain fatty acid synthesis in cell expansion during plant morphogenesis[J].Plant Cell,2005,17(5):1467-1481.
[19]曾艳玲,谭晓风,张党权,等.油茶脂肪酸代谢途径中关键酶基因调控油脂合成的规律研究[J].中国粮油学报,2014,29(2):26-29,35.
[20]Chen G,Peng Z Y,Shan L,et al.Functional expression analysis of an acy-ACP thioesterase Fat B1 from Arachis hypogaea L.seed in Escherichia coli[J].Journal of food,Agriculture&Environment,2012,10(3):332-336.