[关键词]
[摘要]
目的 对龙骨马尾杉Phlegmariurus carinatus法呢二磷酸合成酶(Farnesyl diphosphate synthase,FPS)基因PcFPS1编码区进行克隆及序列分析。方法 根据已获得的龙骨马尾杉转录组数据,从中获得1条编码FPS的转录本,采用RT-PCR方法获得PcFPS1的编码区序列,并对PcFPS1蛋白进行理化性质、蛋白二级结构及三维结构预测分析。结果 序列分析表明,所克隆的PcFPS1基因编码区长为1 119 bp,编码372个氨基酸残基,PcFPS1与挪威云杉Picea abies的FPS具有70%的序列相似性。生物信息学预测PcFPS1蛋白基本不含跨膜区,具有萜类合酶保守结构域,不含信号肽。结论 首次获得龙骨马尾杉PcFPS1基因的编码区序列,为进一步研究PcFPS1在石杉科植物萜类及甾醇类化合物生物合成途径中的功能及鉴定酶活性位点奠定基础。
[Key word]
[Abstract]
Objective To clone and analyze the coding region of Farnesyl diphosphate synthase (FPS) gene PcFPS1 from Phlegmariurus carinatus. Methods According to the acquired transcriptome dataset of P. carinatus, one transcript coding FPS was obtained. The coding region sequence was obtained using RT-PCR method, and the physicochemical properties, protein secondary structure, and three-dimensional structure of PcFPS1 protein were predictively analyzed. Results The cloned PcFPS1 gene contained a 1 119-bp coding region for encoding a predicted protein of 372 amino acids with high homology (70%) to FPS gene in Picea abies. PcFPS1 contained almost no transmembrane region and had the conserved domain of terpenoid cylases, without signal peptide. Conclusion This study clones and analyzes the FPS gene from P. carinatus which is obtained for the first time. The results will provide a foundation for exploring the function of PcFPS1 in terpene and sterol biosynthesis of plants in Huperziaceae.
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[基金项目]
国家自然科学基金项目“基于宏量ESTs的蛇足石杉转录组分析及石杉碱甲合成酶(HAS)的鉴定研究”(30900113);2011年度教育部“长江学者和创新团队发展计划”(IRT1150)