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银杏法呢基二磷酸合酶编码cDNA的克隆及功能分析

Cloning and functional analysis of a cDNA encoding Ginkgo biloba farnesyl diphosphate synthase.

作者信息

Wang Peng, Liao Zhihua, Guo Liang, Li Wenchao, Chen Min, Pi Yan, Gong Yifu, Sun Xiaofen, Tang Kexuan

机构信息

State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, Morgan-Tan International Center for Life Sciences, Fudan University, Shanghai 20433, China.

出版信息

Mol Cells. 2004 Oct 31;18(2):150-6.

Abstract

Farnesyl diphosphate synthase (FPS; EC2.5.1.1/EC2. 5.1.10) catalyzes the synthesis of farnesyl diphosphate, and provides precursor for biosynthesis of sesquiterpene and isoprenoids containing more than 15 isoprene units in Ginkgo biloba. Here we report the cloning, characterization and functional analysis of a new cDNA encoding FPS from G. biloba. The full-length cDNA (designated GbFPS) had 1731 bp with an open reading frame of 1170 bp encoding a polypeptide of 390 amino acids. The deduced GbFPS was similar to other known FPSs and contained all the conserved regions of trans-prenyl chain-elongating enzymes. Structural modeling showed that GbFPS had the typical structure of FPS, the most prominent feature of which is the arrangement of 13 core helices around a large central cavity. Southern blot analysis revealed a small FPS gene family in G. biloba. Expression analysis indicated that GbFPS expression was high in roots and leaves, and low in stems. Functional complementation of GbFPS in an FPS-deficient strain confirmed that GbFPS mediates farnesyl diphosphate biosynthesis.

摘要

法尼基二磷酸合酶(FPS;EC2.5.1.1/EC2.5.1.10)催化法尼基二磷酸的合成,并为银杏中倍半萜和含有超过15个异戊二烯单元的类异戊二烯的生物合成提供前体。在此,我们报道了从银杏中克隆、鉴定和功能分析一个编码FPS的新cDNA。全长cDNA(命名为GbFPS)有1731 bp,开放阅读框为1170 bp,编码一个390个氨基酸的多肽。推导的GbFPS与其他已知的FPS相似,并包含反式异戊二烯链延长酶的所有保守区域。结构建模表明,GbFPS具有FPS的典型结构,其最显著的特征是围绕一个大的中央腔排列着13个核心螺旋。Southern杂交分析揭示了银杏中有一个小的FPS基因家族。表达分析表明,GbFPS在根和叶中表达较高,在茎中表达较低。在一个FPS缺陷菌株中对GbFPS进行功能互补,证实GbFPS介导法尼基二磷酸的生物合成。

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