Cloning and functional analysis of farnesyl pyrophosphate synthase (FPPS) gene fromMylabris cichorii

焦磷酸法尼酯 斑蝥素 基因 生物 基因表达 克隆(编程) 同源(生物学) ATP合酶 生物化学 分子生物学 遗传学
作者
Shenfang Zha,Youping Yin,Yu Wang,Yi Huang,Yan Li,Zhongkang Wang
出处
期刊:Biotechnology and Applied Biochemistry [Wiley]
卷期号:64 (5): 667-676 被引量:12
标识
DOI:10.1002/bab.1494
摘要

Cantharidin, a defensive terpene compound synthesized by the meloid beetle (Coleoptera, Meloidae), is an important anticancer agent. However, there has been little study done on how this compound synthesized by the beetle. In this paper, a farnesyl pyrophosphate synthase (FPPS) gene, designated McFPPS, was isolated from Mylabris cichorii by reverse transcription PCR based on conserved domains in other organisms. Multiple alignment analysis showed that the deduced amino acids shared >70% homology with FPPSs from other species and contained typically seven conservative regions. Gene expression profile analysis revealed that McFPPS was expressed throughout the tested growth stages of M. cichorii adults, whereas the transcripts accumulated to the highest level at 20 days in male adults while the highest expression level appeared at 15 days in females. Tissue expression pattern analysis showed that McFPPS was expressed constitutively in all tested tissues and a relatively higher expression level in the alimentary canal of males, but no significant tissue difference in the females. For the first time, a RNA interference strategy was employed to induce a greater suppression of McFPPS mRNA, and thus a sharp decrease in the expression levels of downstream genes and the concentration of product. All these results indicated that McFPPS may be directly involved or play an essential role in the biosynthesis of cantharidin.

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