目的 基于前期对白木香Aquilaria sinensis转录组测序结果,从白木香总RNA中克隆白木香倍半萜合成酶基因(As-SesTPS1),并对其进行生物信息学及表达分析。方法 利用逆转录聚合酶链式反应(RT-PCR)从白木香总RNA中获得倍半萜合成酶基因As-SesTPS1的cDNA全长序列;利用生物信息学手段对该序列进行相似性和同源性分析,并预测其编码蛋白的结构和功能;利用qRT-PCR技术检测该基因在白木香不同样品中的表达水平。结果 得到倍半萜合成酶基因As-SesTPS1,开放阅读框(ORF)全长1 671 bp,编码556个氨基酸,其与多条倍半萜合成酶基因具有较高相似性,并含有RRx₈W和DDxxD的保守序列;该基因在白木香的沉香样品中高表达。结论 As-SesTPS1的克隆及其生物信息学和在白木香不同部位表达差异性分析,为白木香倍半萜生物合成代谢途径的研究奠定基础。

Objective To clone a sesquiterpene synthase gene denoted As-SesTPS1 from total RNA of Aquilaria sinensis based on previous transcriptome sequencing data and analyze the bioinformatics and expression of the gene. Methods The full length cDNA of As-SesTPS1 was obtained by reverse transcription-PCR (RT-PCR). The sequence similarity and homology were analyzed, and the structure and function of the coding protein were predicted by bioinformatics method. The gene expression levels in different samples of A. sinensis were quantified using qRT-PCR technique. Results The sesquiterpene synthase gene As-SesTPS1 was cloned, which contained an ORF of 1 671 bp, encoding 556 amino acids, and shared high similarity to multiple sesquiterpene synthase genes. The protein encoded by this gene had conserved RRx₈W and DDxxD motifs. It was confirmed the gene was highly expressed in agarwood sample through qRT-PCR technique. Conclusion The sesquiterpene synthase gene As-SesTPS1 is cloned, which will lay a foundation for further study of sesquiterpene biosynthase pathway in A. sinensis.

国家自然科学基金项目(31100496,81102418);"973"前期专项(2014CB460613);广东省科技计划项目(2012A030100014);广东省中国科学院全面战略合作项目(2011B090300078);广东省科学院野外科学实验站基金项目(Sytz201204)