• 中文核心期刊
  • CSCD来源期刊
  • 中国科技核心期刊
  • CA、CABI、ZR收录期刊

茉莉花JsMYB305转录因子互作蛋白的筛选及验证

Interacting Proteins with JsMYB305 Transcription Factor in Jasminum sambac

  • 摘要:
      目的  茉莉花JsMYB305是参与萜烯合成酶(Terpene synthetases,TPS)基因调控茉莉花香气的转录因子。本研究旨在进一步分析JsMYB305转录因子调控茉莉花香气物质代谢的可能机制。
      方法  基于茉莉花瓣酵母文库,以白色花苞和完全开放时的茉莉花为材料构建双杂交文库,通过酵母双杂交的方式筛选JsMYB305的互作蛋白并进行鉴定和验证。
      结果  JsMYB305全长具有显著的自激活性,分别扩增不同长度的片段进行自激活验证,当编码长度≤510 bp时,JsMYB305没有自激活性。进一步以pGBKT7-JsMYB305(510 bp)为诱饵,从酵母双杂交文库中筛选到1个互作蛋白,经鉴定为水杨酸甲酯转移酶(Salicylic acid carboxyl methyltransferase,SAMT)蛋白。酵母定向双杂验证表明JsMYB305与JsSAMT具有相互作用。
      结论  SAMT属于苯丙烷/丙环生物合成途径,故JsMYB305转录因子也参与了茉莉花苯丙烷/丙环类香气物质合成过程。研究结果为进一步深入研究JsMYB305在茉莉花香气合成中的调控机理提供了参考。

     

    Abstract:
      Objective   Aroma-regulating mechanism of the transcription factor JsMYB305 associated with terpene synthetases gene of jasmine flower was investigated.
      Method   From the jasmine floral petal yeast database a two-hybrid library was constructed using the white buds and fully open flowers. Proteins that interacted with JsMYB305 were identified and verified by the yeast two-hybrid method.
      Result   The full length of JsMYB305 was significantly self-activated, and the fragments of different lengths were amplified for verification. At a coding length less than or equal to 510 bp, JsMYB305 showed no self-activation. Applying a 510 bp pGBKT7-JsMYB305 as a bait, an interacting protein was obtained from the yeast two-hybrid library and identified to be salicylic acid carboxyl methyltransferase (SAMT). The yeast-directed two-hybrid validation indicated that JsMYB305 interacted with JsSAMT.
      Conclusion   Since SAMT presents in the biosynthetic pathway of phenylpropane/propane, the JsMYB305 transcription factor was believed to also relate to the synthesis of similar aromatic substances in jasmine flowers.

     

/

返回文章
返回