蒺藜苜蓿DGAT1基因的克隆和功能鉴定
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国家自然科学基金青年基金(31300569);


Cloning and Functional Characterization of Diacylglycerol Acyltransferase Gene (MtDGAT1) from Medicago truncatula
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    摘要:

    该研究采用RTPCR与电子克隆的方法,从蒺藜苜蓿cDNA中克隆得到2个编码二脂酰甘油酰基转移酶(diacylglycerol acyltransferase,DGAT)的基因MtDGAT11和MtDGAT12。MtDGAT11长1 620 bp,编码539个氨基酸;MtDGAT12长1 524 bp,编码507个氨基酸。多序列比对显示,MtDGAT11和MtDGAT12编码蛋白具有典型的植物DGAT1结构域。表达分析显示,MtDGAT11和MtDGAT12在根、茎、叶、花、种子中都有表达,在种子发育中高表达,且MtDGAT11于种子发育的中前期高表达,而MtDGAT12于种子发育的中后期高表达。酵母互补实验证实,MtDGAT12编码蛋白具有DGAT酶活性,能够恢复H1246的TAG合成和油体形成;而MtDGAT11编码蛋白不能恢复H1246的TAG合成和油体形成。

    Abstract:

    In this study, we acquired two MtDGAT1 genes (MtDGAT11 and MtDGAT12) from cDNA of Medicago truncatula by using insilico cloning combined with RTPCR. The length of MtDGAT11 is 1 620 bp, encoding 539 amino acids, while the length of MtDGAT12 is 1 524 bp, encoding 507 amino acids. Multiple alignments revealed that both MtDGAT11 and MtDGAT12 shared the typical functional motifs with DGAT1s from other plant species. Expression analysis showed that both MtDGAT11 and MtDGAT12 were expressed in root, stem, leaf, flower and seed, and highly expressed in developing seeds; MtDGAT11 was highly expressed in earlymiddle periods of seed development, while MtDGAT12 was highly expressed in the middle and later periods of seed development. Functional complementation in yeast H1246 confirmed that MtDGAT11 encodes disfunctional DGAT and MtDGAT12 encodes functional DGAT.

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徐荣华,崔 涛,王建才,等.蒺藜苜蓿DGAT1基因的克隆和功能鉴定[J].西北植物学报,2016,36(10):1941-1947

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  • 在线发布日期: 2016-11-14
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