Cloning and functions analysis of a pyruvate dehydrogenase kinase in Brassica napus
Cloning and functions analysis of a pyruvate dehydrogenase kinase in Brassica napus
复制标题
甘蓝型油菜丙酮酸脱氢酶激酶的克隆及功能分析
DOI：
10.1007/s00299-011-1066-2
复制
发表时间：
2011-08-01
影响因子：
6.2
通讯作者：
Hua, Wei
中科院分区：
文献类型：
--
作者：
Li, Rong-Jun;Hu, Zhi-Yong;Hua, Wei
丙酮酸脱氢酶激酶(PDK)是线粒体丙酮酸脱氢酶复合体(MtPDC)的负性调节因子，在中间代谢中起关键作用。本研究从甘蓝型油菜(Brassica napus，BnPDK1)基因文库中分离和克隆了1,490bpPDK基因。BnPDK1有1,104个开放阅读框，编码367个氨基酸。基因组DNA凝胶印迹分析结果表明，BnPDK1是一个多拷贝基因。用RNA凝胶印迹分析和RNA原位杂交检测BnPDK1在不同器官中的表达。BnPDK1基因在几乎所有供试组织中都有表达，其中以雄蕊和幼角果的表达最高。BnPDK1在转基因拟南芥中的过表达会抑制PDC活性，导致种子含油量和叶片光合作用下降。这些结果表明，BnPDK1参与了种子发育过程中脂肪酸生物合成的调控。
Pyruvate dehydrogenase kinase (PDK) is a negative regulator of the mitochondrial pyruvate dehydrogenase complex (mtPDC), which plays a key role in intermediary metabolism. In this study, a 1,490-bp PDK in Brassica napus (BnPDK1) was isolated and cloned from Brassica cDNA library. BnPDK1 has an 1,104 open reading frame encoding 367 amino acids. Genomic DNA gel blot analysis result indicated that BnPDK1 is a multi-copy gene. RNA gel blot analysis and RNA in situ hybridization were used to determine the expression of BnPDK1 in different organs. BnPDK1 gene was ubiquitously expressed in almost all the tissues tested, having the highest expression in the stamen and the young silique. Over-expression of BnPDK1 in transgenic Arabidopsis lines would repress the PDC activity, and resulted in the decrease of seed oil content and leaf photosynthesis. These results implied that BnPDK1 was involved in the regulation of fatty acid biosynthesis in developing seeds.