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以双瓣茉莉花[Jasminum sambac(L.)Ait]花瓣为材料,采用RT-PCR和RACE技术相结合的方法,克隆了萜类合成酶基因(Js TPS)的全长cDNA,该cDNA全长为1 884 bp,其中ORF为1 491 bp,编码496个氨基酸的蛋白,分子量56 989.7 D,与原核表达结果一致。序列分析结果表明,该基因编码的氨基酸序列与油橄榄(Olea europaea)的TPS2具有75%的同源性,同属于α-Farnesene synthase分支。采用荧光定量PCR技术检测Js TPS在茉莉花开放过程中的表达量变化,结果表明,表达量在未开放时(18:00)最低,在半开放时(22:00)达到最高。
The full-length cDNA of terpenoid synthase gene (Js TPS) was cloned by RT-PCR and RACE techniques using the petals of Jasminum sambac (L.) Ait. 1 884 bp. The ORF was 1 491 bp, encoding a protein of 496 amino acids with a molecular mass of 56 989.7 D, which was consistent with the result of prokaryotic expression. Sequence analysis showed that the deduced amino acid sequence was 75% homologous to TPS2 of Olea europaea, belonging to the α-Farnesene synthase branch. Fluorescence quantitative PCR was used to detect the expression of Js TPS during the opening of jasmine. The results showed that the expression of Js TPS was the lowest at 18:00 and reached the highest at 22:00.