福建农业学报2025，Vol.40Issue(12)：1232-1239,8.DOI:10.19303/j.issn.1008-0384.2025.12.002
红麻植株酵母文库的构建及其转录因子HcWRKY71互作蛋白的筛选
Yeast Hybrid Library and HcWRKY71 Interacting Proteins of Kenaf
摘要
Abstract
[Objective]Regulatory network of HcWRKY71 in response to salt stress in kenaf was studied,and proteins interacting with the transcription factor identified.[Method]Roots,stems,leaves,and phloem of Hibiscus cannabinus L.were used to construct a yeast two-hybrid cDNA library using the Gateway method. Proteins interacting with HcWRKY71 were screened,sanger-sequenced,and validated by means of one-to-one rotation.[Result]The constructed yeast library had a capacity of 1.2×107 CFU,a recombination rate of 100%,and an average insertion fragment length of over 1,000bp. Screened by the yeast two-hybrid,48 positive clones interacting with the transcription factor HcWRKY71 were identified. After one-to-one interaction verification,the MYB transcription factor 81,triple helix transcription factor ASIL2,and BAHD acyltransferase protein may participate in the response process of interacting proteins related to kenaf salt stress by interacting with HcWRKY71.[Conclusion]The large-capacity,wide-coverage,and high-quality yeast hybrid library for kenaf established by this study could become a useful tool for research. The selection of potential proteins that might interact with the transcription factor HcWRKY71 would aid future studies in elucidating the molecular mechanism of the plant under salt stress.关键词
红麻/酵母文库/酵母双杂交/WRKY转录因子/盐胁迫Key words
Hibiscus cannabinus L./yeast library/yeast two-hybrid library/WRKY transcription factor/salt stress分类
农业科技引用本文复制引用
李辉,唐慧娟,唐映红,栾明宝..红麻植株酵母文库的构建及其转录因子HcWRKY71互作蛋白的筛选[J].福建农业学报,2025,40(12):1232-1239,8.基金项目
湖南省自然科学基金面上项目(2024JJ5290) （2024JJ5290）
国家现代农业产业技术体系建设项目(CARS-16-E5) （CARS-16-E5）
农业农村部麻类生物学与加工重点实验室开放课题(LBFC-202503) （LBFC-202503）
