生物技术通报 ›› 2021, Vol. 37 ›› Issue (7): 137-145.doi: 10.13560/j.cnki.biotech.bull.1985.2021-0010

• 研究报告 • 上一篇    下一篇

淡紫紫孢菌亲环蛋白PlCYP6 互作蛋白的筛选

刘娟(), 朱春晓, 肖雪琼, 莫陈汨, 王高峰(), 肖炎农()   

  1. 华中农业大学植物科学技术学院,武汉 430070
  • 收稿日期:2021-01-04 出版日期:2021-07-26 发布日期:2021-08-13
  • 作者简介:刘娟,女,硕士研究生,研究方向:分子植物病理学;E-mail: juandalin0704@163.com
  • 基金资助:
    国家自然科学基金项目(31872019);江西省重点研发计划项目(20181ACF60018)

Screening of Protein Interacting with Purpureocillium lilacinum Cyclophilin PlCYP6

LIU Juan(), ZHU Chun-xiao, XIAO Xue-qiong, MO Chen-mi, WANG Gao-feng(), XIAO Yan-nong()   

  1. College of Plant Science and Technology,Huazhong Agricultural University,Wuhan 430070
  • Received:2021-01-04 Published:2021-07-26 Online:2021-08-13

摘要:

以淡紫紫孢菌中参与盐胁迫响应的亲环蛋白PlCYP6 为研究对象,采用免疫沉淀联合质谱分析及酵母双杂交等技术筛选淡紫紫孢菌中与PlCYP6 互作的蛋白。结果显示,受PlCYP6 特异钓取的482 个蛋白的功能主要涉及细胞代谢。其中,乙醇脱氢酶1(alcohol dehydrogenase I,ADH1)与PlCYP6 直接互作,且PlCYP6 的WD40 repeat 结构域为二者间互作的关键区域。同时,PlCYP6 和ADH1 均受NaCl 胁迫诱导表达。上述研究结果表明,ADH1 为PlCYP6 的候选互作蛋白,这为进一步解析淡紫紫孢菌响应盐胁迫机制奠定了基础。

关键词: 淡紫紫孢菌, PlCYP6, 盐胁迫应激反应, 免疫沉淀联合质谱, 互作蛋白

Abstract:

The cyclophilin PlCYP6 that is involved in the responses of Purpureocillium lilacinum to salt stresses was used as the research object,and the putative interaction protein of PlCYP6 in P. lilacinum was screened using immunoprecipitation combined with mass spectrometry(IP-MS),yeast two-hybrid assay and other techniques. Results indicated that 482 proteins was specially fished by PlCYP6,and mainly functioned in cell metabolism. Among them,alcohol dehydrogenase 1(ADH1)directly interacted with PlCYP6,and the domain,WD40 repeat,of PlCYP6 was involved the interaction between them. Meanwhile,both PlCYP6 and ADH1 were induced by NaCl stress. These findings reveal that ADH1 is a putative interaction protein of PlCYP6,which lays a foundation for further uncovering the mechanism of responses of P. lilacinum to salt stresses.

Key words: Purpureocillium lilacinum, PlCYP6, salt stress response, immunoprecipitation combined with mass spectrometry(IP-MS), interaction protein