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甘蓝型油菜丙酮酸脱氢酶激酶基因的克隆与功能分析。

Cloning and functions analysis of a pyruvate dehydrogenase kinase in Brassica napus.

机构信息

Department of Chemistry, Wuhan University, Wuhan, 430072, People's Republic of China.

出版信息

Plant Cell Rep. 2011 Aug;30(8):1533-40. doi: 10.1007/s00299-011-1066-2. Epub 2011 Apr 2.

Abstract

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.

摘要

丙酮酸脱氢酶激酶(PDK)是线粒体丙酮酸脱氢酶复合物(mtPDC)的负调节剂,在中间代谢中起着关键作用。本研究从油菜 cDNA 文库中分离并克隆了一个 1490bp 的 PDK(BnPDK1)。BnPDK1 具有一个 1104bp 的开放阅读框,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。RNA 凝胶印迹分析和 RNA 原位杂交用于确定 BnPDK1 在不同器官中的表达。BnPDK1 基因在几乎所有测试的组织中都有广泛表达,在雄蕊和幼角果中表达最高。在转基因拟南芥系中过表达 BnPDK1 会抑制 PDC 活性,导致种子油含量和叶片光合作用降低。这些结果表明 BnPDK1 参与了发育种子中脂肪酸生物合成的调节。

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