油茶CoPDAT基因的克隆与表达分析

发布时间：2018-07-09 18:05

  本文选题：基因克隆 + 生物信息学分析　； 参考：《植物生理学报》2017年09期

【摘要】：磷脂:二酰甘油酰基转移酶(PDAT)能够催化磷脂酰胆碱(PC)sn-2位的酰基转移至二酰甘油(DAG)上,形成三酰甘油(TAG)和溶血磷脂酰胆碱,这是另外一条不同于Kennedy途径的三酰甘油合成方式,故明确油茶(Camellia oleifera)PDAT基因的功能对阐明油茶种子油脂合成机制具有重要意义。本研究通过cDNA末端快速扩增(RACE)技术克隆油茶CoPDAT基因的全长cDNA序列,该序列长2773 bp,开放阅读框为2022 bp,编码673个氨基酸。亚细胞定位分析确定CoPDAT基因所编码蛋白质定位于内质网上。实时荧光定量PCR分析结果表明,CoPDAT在油茶不同组织/器官中均有表达,在种子中不同发育时期的相对表达量呈"上升-下降-上升-下降-上升"的表达特征,在花后42周表达量最高。CoPDAT基因与油茶种子油脂含量的灰色关联度值为0.8356,表明该基因与油茶种子油脂的合成积累关系密切。本研究为阐明CoPDAT基因在油茶油脂合成过程中的功能以及为油茶的分子遗传改良提供参考。
[Abstract]:Phospholipid: diacylglycerol transferase (PDAT) catalyzes the transfer of acyl groups from the sn-2 site of phosphatidylcholine (PC) to diacylglycerol (DAG) to form triglycerides (tag) and lysophosphatidylcholine (lysophosphatidylcholine). Therefore, it is important to clarify the function of Camellia oleifera PDAT gene in oil synthesis mechanism of Camellia oleifera. In this study, the full-length cDNA sequence of Camellia oleifera CoPDAT gene was cloned by rapid amplification of cDNA terminal (race). The length of the cDNA was 2773 BP, and the open reading frame was 2022 BP, encoding 673 amino acids. Subcellular localization analysis confirmed that the protein encoded by CoPDAT gene was located on the endoplasmic reticulum. The results of real-time fluorescence quantitative PCR analysis showed that CoPDAT was expressed in different tissues / organs of Camellia oleifera, and the relative expression level of CoPDAT in different development stages of Camellia oleifera was "upper-down-ascendant". At 42 weeks after anthesis, the highest expression of .CoPDAT gene and the grey-related degree of oil content of Camellia oleifera seeds were 0.8356, which indicated that this gene was closely related to the synthesis and accumulation of oil in Camellia oleifera seeds. This study was intended to elucidate the function of CoPDAT gene in the synthesis of Camellia oleifera oil and provide a reference for the molecular genetic improvement of Camellia oleifera.
【作者单位】： 中南林业科技大学经济林培育与保护教育部重点实验室经济林育种与栽培国家林业局重点实验室经济林培育与利用湖南省2011协同创新中心;
【基金】：国家自然科学基金(31470684)~~
【分类号】：Q943.2;S794.4
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本文编号：2110183