牛樟芝萜烯合成酶基因的克隆与鉴定

为了研究牛樟芝(Antrodia camphorata)倍半萜的生物合成,通过对牛樟芝基因组分析获得倍半萜类合成酶基因(AcTPS2),利用RT-PCR克隆获得其全长cDNA,对其进行生物信息学分析和表达谱分析。结果显示:AcTPS2基因cDNA全长1 068bp,Blast比对发现,AcTPS2含倍半萜类合成酶所独具的富含天冬氨酸序列DXXXD及萜类合成酶特有的RRDTSG-LDL保守序列;系统进化分析显示,AcTPS2基因与其他真菌的倍半萜聚为一类;表达谱分析显示,以甘露糖作为碳源、以酪蛋白胨作为氮源能够有效促进AcTPS2基因的诱导表达。研究结果可为以后牛樟芝倍半萜类生物合成提供一定的参考依据。

Cloning and Identification of Terpene Synthetases Gene in Antrodia camphorata

In order to study the biosynthesis of sesquiterpene, this text analyzed the sesquiterpene synthetases gene(AcTPS2) through genome analysis of Antrodia camphorata, and obtained the full length cDNA with RT PCR, carried on bioinformatics and mRNA analysis. The results showed that AcTPS2 cDNA has 1 086 bp; Blast analysis revealed that AcTPS2 included abundant asparagic acid motif DXXXD that is specific motif of sesquiterpene synthetase, and terpene synthetase special conserved sequence RRDTSG LDL; phylogenetic trees showed AcTPS2 and other fungus sesquiterpene synthetase amino acid sequence clustered in one clade; mRNA analysis showed manitol as carbon source, casein peptone as nitrogen source could promote AcTPS2 effectively, and the results could supply some basis in sesquiterpene biosynthesis.