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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。
Objective To clone and sequence the coding region of PcFPS1 in Phlegmariurus carinatus, a Farnesyl diphosphate synthase (FPS) gene. Methods According to the obtained transcriptome data of Horus johnii, one transcript of FPS was obtained and the coding region of PcFPS1 was obtained by RT-PCR. The physicochemical properties, protein secondary structure and three-dimensional structure of PcFPS1 protein were predicted analysis. Sequence analysis showed that the cloned PcFPS1 gene was 1 119 bp in length and encoded 372 amino acid residues. The sequence similarity of PcFPS1 with FPS of Picea abies in Norway was 70%. Bioinformatics prediction PcFPS1 protein is basically free of transmembrane region, with terpene conservative conserved domain, without signal peptide. Conclusions The coding region of PcFPS1 gene was obtained for the first time in order to further study the function of PcFPS1 in the biosynthesis pathway of terpenoids and sterols in the genus Pinaceae and to lay a foundation for the identification of the enzyme active sites.