蛇足石杉鲨烯合酶HsSQS1基因克隆和序列分析

Cloning and analysis of squalene synthase(HsSQS1) gene in Huperzia serrata

鲨烯合酶(squalene synthase,SQS)是植物萜类化合物生物合成途径的关键酶。本研究根据课题组已获得的蛇足石杉(Huperzia serrata)转录组数据中的SQS1转录本序列,设计基因全长扩增引物,利用RT-PCR方法获得蛇足石杉SQS1基因的全长cDNA序列;利用实时荧光定量PCR方法检测了HsSQS1基因在蛇足石杉根、茎、叶中的表达情况;对HsSQS1基因进行生物信息学分析,并预测HsSQS1编码蛋白的结构与功能。研究结果表明,克隆获得的蛇足石杉SQS1基因长为1 263 bp,编码420个氨基酸,命名为HsSQS1,GenBank登录号JQ004938。HsSQS1基因在蛇足石杉的根中表达丰度高于茎和叶。本研究成功克隆并分析了蛇足石杉SQS1的全长序列,为进一步阐明蛇足石杉三萜代谢途径奠定基础。

Squalene synthase（SQS） is a key enzyme in plant terpenoid biosynthetic pathway.This study focused on cloning and analysis of Huperzia serrata SQS（HsSQS1） gene.After searching the transcriptome dataset of H.serrata,one unique sequence encoding SQS was discovered.The primers were designed according to the transcript sequence of HsSQS1 from the H.serrata transcriptome dataset.The open reading frame of HsSQS1 was cloned using RT-PCR strategy.The bioinformatic analysis of this gene and its corresponding protein were performed.The cDNA（named as HsSQS1） contains a 1 263 bp open reading frame and encodes a predicted protein of 420 amino acids.The GenBank accession number for this gene is JQ004938.HsSQS1 contains two transmembrane regions,without signal peptide.The conserved domain of squalene synthase was presented in HsSQS1.HsSQS1 was more abundant in H.serrata root than in leaf and stem.This study cloned and analyzed squalene synthase gene from H.serrata for the first time.The result will provide a foundation for exploring the mechanism of terpenoid biosynthesis in H.serrata plants.