河南农业大学学报2024，Vol.58Issue(2)：218-227,10.DOI:10.16445/j.cnki.1000-2340.20240030.001
麦根腐平脐蠕孢cDNA文库的构建及BsTup1互作蛋白筛选
Construction of cDNA library of Bipolaris sorokiniana and screening of BsTup1 interacting proteins
摘要
Abstract
[Objective]To investigate the molecular mechanisms of growth,development,and pathogenicity of Bipolaris sorokiniana and identify the interacting proteins of BsTup1.[Method]We used spores and mycelia at different stages of B.sorokiniana as materials to construct a yeast two-hybrid cDNA library,and screened the yeast two-hybrid library by BsTup1 gene as the bait for determining the proteins interacting with BsTup1.[Result]1)The mixed full-length cDNA library of conidia and mycelium was successfully constructed for the first time using SMART(switching mechanism at 5'end of the RNA transcript)technology.The results of library identification showed that the constructed cDNA library had a capacity of 4.8×107 cfu·mL-1,the library insertion fragment recombination rate reached 100％and the average size was around 1 000 bp.2)The bait protein vector pGBKT7-BsTup1,a transcriptional inhibitor of BsTupl in B.sorokiniana,successfully constructed and confirmed no self-activation activity.3)Using this bait vector,38 candidate proteins interacting with BsTup1 were obtained after sequencing,sequence alignment,and yeast verification.[Conclusion]We successfully constructed a cDNA library of B.sorokiniana,and identified 38 candidate proteins which interacted with BsTup1.关键词
麦根腐平脐蠕孢/SMART技术/cDNA文库/酵母双杂/BsTup1Key words
Bipolaris sorokiniana/SMART technology/cDNA library/yeast two-hybrid/BsTup1分类
农业科技引用本文复制引用
李长水,耿月华,姚萌,赵炳森,谢顺培,徐超,马庆周,张猛..麦根腐平脐蠕孢cDNA文库的构建及BsTup1互作蛋白筛选[J].河南农业大学学报,2024,58(2):218-227,10.基金项目
中国博士后基金第74批面上资助项目(2023M741063) （2023M741063）
河南省科技攻关项目(232102110073) （232102110073）
河南省高校科技创新团队支持计划项目(18IRTSTHN021) （18IRTSTHN021）
河南农业大学高层次人才专项支持基金项目(30501502) （30501502）
河南农业大学作物学科合作交叉优中选优项目(CCSR2022-3) （CCSR2022-3）
