牛樟芝羊毛甾醇合成酶基因AcLSS的克隆和表达分析

Cloning and Expression Analysis of the Lanosterol Synthetase Gene(AcLSS)from Antrodia cinnamomea

为探究牛樟芝羊毛甾醇合成酶的合成机制,对牛樟芝的一个羊毛甾醇合成酶基因进行克隆,将其命名为AcLSS,然后利用生物信息学软件对其理化性质进行预测,并对其表达情况进行初步分析。结果表明:AcLSS基因的开放阅读框为2205 bp,编码734个氨基酸;该基因含有8个内含子,9个外显子;AcLSS蛋白存在由14个氨基酸“KGAWPFSTKTQGYT”和6个氨基酸“VSDCTG”组成的保守特征序列,并且包含一个16个氨基酸组成的OSC保守结合域“KACNFLISKQRSDGGW”。分子系统进化分析显示,AcLSS蛋白与绣球菌(XP_027617410.1)的LSS蛋白亲缘关系最近;表达谱分析显示,碳源添加物中以甘露醇的诱导表达力最高、氮源添加物中以番茄浸粉的诱导表达力最佳。

In order to study the biosynthesis mechanism,a lanosterol synthase gene(AcLSS)from Antrodia cinnamomea was isolated and cloned in this study,and then the physicochemical properties were predicted by using bioinformatics software,and the gene expressions were also analyzed.The results showed that the open reading frame of AcLSS gene was 2205 bp,which encoding 734 amino acids;AcLSS gene had 8 introns and 9 exons;AcLSS protein had conserved characteristic sequences which consist of 14 amino acids'KGAWPFSTKTQGYT'and 6 amino acids'VSDCTG',as well as a OSC conserved domain which consist of 16 amino acids'KACNFLISKQRSDGGW'.The molecular phylogenetic analysis revealed that AcLSS protein was most closely related to the LSS protein of Sparassis crispa(XP_027617410.1).The gene expression profile indicated that the best ability of induction expression AcLSS gene was the mannitol from carbon source and the tomato powder from nitrogen source,respectively.