生物技术通报2024，Vol.40Issue(6)：281-289,9.DOI:10.13560/j.cnki.biotech.bull.1985.2024-0121
玉米大斑病菌cDNA文库的构建及转录因子StMR1互作蛋白的筛选
Construction of cDNA Library of Setosphaeria turcica and Screening of Transcription Factor StMR1 Interacting Proteins
摘要
Abstract
[Objective]To screen the interaction proteins of transcription factors of Setosphaeria turcica,and to analyze the molecular mechanism of melanin-regulated transcription factor StMR1 in regulating the pathogenicity of S.turcica.This study provides a reference for elucidating the regulatory network of transcription factors in the infection process of S.turcica and elucidating the pathogenic mechanism of the pathogen.[Method]Different germination stages of hyphae and spores of S.turcica were collected as test materials,and the cDNA library of S.turcica was constructed by gateway method,The bait vector of transcription factor StMR1 was constructed by homologous recombination,and its interacting proteins were screened by yeast two-hybrid technology and verified one-to-one.[Result]The average fragment length of the constructed library was greater than 1 000 bp,the library capacity of the primary library and the secondary library were 1.2×107 and 1.04×107 CFU,respectively,and the recombination rate was 100%,which could be used for yeast two-hybrid screening.The bait vector pGBKT7-StMR1 that could be used for screen library was successfully constructed,and three interacting proteins were obtained through primary screening and re-screening,and one-to-one verification of the interaction between short-chain dehydrogenase,glycosyltransferase and leucine-rich repeat protein and transcription factor StMR1 were verified.[Conclusion]A high quality and abundant cDNA library was successfully constructed and the protein interacting with StMR1 was screened关键词
玉米大斑病菌/cDNA文库/转录因子/酵母双杂交/互作蛋白Key words
Setosphaeria turcica/cDNA library/transcription factor/yeast two-hybrid/interaction protein引用本文复制引用
王秋月,段鹏亮,李海笑,刘宁,曹志艳,董金皋..玉米大斑病菌cDNA文库的构建及转录因子StMR1互作蛋白的筛选[J].生物技术通报,2024,40(6):281-289,9.基金项目
国家自然科学基金项目(32072370),河北省自然科学基金项目(C2021204136),国家现代农业产业技术体系(CARS-02) （32072370）
