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园艺学报 ›› 2016, Vol. 43 ›› Issue (2): 356-364.doi: 10.16420/j.issn.0513-353x.2015-0803

• 研究报告 • 上一篇    下一篇

茉莉花萜类合成酶基因JsTPS的克隆及其表达分析

俞 滢1,*,陈 丹1,*,孙 君2,吕恃衡1,陈桂信1,**,叶乃兴1,**   

  1. 1福建农林大学园艺学院/茶学福建省高等学校重点实验室,福州 350002;2福建省农业科学院茶叶研究所,福建福安 355015
  • 出版日期:2016-02-25 发布日期:2016-02-25
  • 基金资助:
    福建省自然科学基金项目(2016J01110);福州市科技局市校合作项目(2013-G-103);福州市农业局2014年福州茉莉花茶产业提升项目(2014-3)

Cloning and Expression Analysis of Terpene Synthase Gene from Jasminum sambac

YU Ying1,*,CHEN Dan1,*,SUN Jun2,Lü Shi-heng1,CHEN Gui-xin1,**,and YE Nai-xing1,**   

  1. 1College of Horticulture,Fujian Agriculture and Forestry University,Key Laboratory of Tea Science at Universities in Fujian,Fuzhou 350002,China;2Tea Research Institute,Fujian Academy of Agricultural Sciences,Fu’an,Fujian 355015,China
  • Online:2016-02-25 Published:2016-02-25

摘要: 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(JsTPS)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JsTPS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。

关键词: 茉莉花, 萜类合成酶, 实时荧光定量PCR, 原核表达

Abstract: The full length cDNA of terpene synthase gene(JsTPS)was cloned by combination of RT-PCR and RACE from petals of Jasminum sambac. The results showed that full length cDNA contained 1 491 bp including an 1 884 bp ORF,encoding a 56 989.7 D protein with 496 amino acids whose molecular weight was consistent to that of product of the gene by prokaryotic expression;The result of alignment of amino acid showed that the gene had a homology of 75% to that of olive(Olea europaea),belonging to α-Farnesene synthases;Quantities of the gene were detected by real-time PCR in process of opening of flower,the results showed that the expressing level of the gene was minimum at 18:00 when the flowers were not open,then had been increasing until at 22:00 when the expressing level reached to maximum and the flowers was opening.

Key words: Jasminum sambac, terpene synthase gene, quantitative real-time PCR, prokaryotic expression

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