棒曲霉孢子发育相关基因flbA的克隆和缺失突变株的鉴定  

Cloning of flbA gene related to conidial development of Aspergillus clavatus and identification of deletion mutants

作　　者：蒋冬花 吕梦霞 史婷婷 丁允章 许楚旋 JIANG Donghua;LYU Mengxia;SHI Tingting;DING Yunzhang;XU Chuxuan(College of Chemistry and Life Sciences,Zhejiang Normal University,Jinhua 321004,China)

机构地区：[1]浙江师范大学化学与生命科学学院

出　　处：《浙江师范大学学报（自然科学版）》2020年第1期1-7,共7页Journal of Zhejiang Normal University:Natural Sciences

基　　金：国家自然科学基金资助项目(31570013);浙江省公益技术研究计划资助项目(GN18C010004)

摘　　要：在模式真菌构巢曲霉中,FlbA蛋白位于分生孢子中心调控途径上游,对分生孢子的形成起激活作用.研究以1株实验室筛选的高产洛伐他汀棒曲霉Ac-32专利菌株的基因组为模板,克隆分生孢子发育相关基因flbA,用双接头PCR法构建flbA基因敲除盒,用根癌农杆菌介导转化法构建棒曲霉转化子库,依据转化子表型特征和分子特征鉴定flbA基因缺失突变株.结果表明:flbA基因全长为2208 bp,翻译对应的氨基酸为734个,理论分子量和等电点分别为78798.75 Da和8.64;棒曲霉与构巢曲霉flbA基因序列同源性为69.9%,氨基酸序列的同源性为76.6%;构建了flbA基因敲除盒和敲除载体,获得了258株棒曲霉转化子,经鉴定得到了1株flbA基因缺失突变株.研究成果为后续探讨棒曲霉flbA基因功能提供了材料,并奠定了理论基础.In model fungi Aspergillus nidulans,FlbA protein located in the upstream of the conidial central regulatory pathway activated the formation of conidia.The conidial development-related flbA gene was cloned from a laboratory-screened patent strain of Lovastatin-producing A.clavatus Ac-32.The flbA gene knockout box was constructed by double-junction PCR,and the transformant library of A.clavatus was constructed by Agrobacterium tumefaciens-mediated transformation.The results showed that the total length of flbA gene was 2208 bp,and the translated amino acids were 734.The theoretical molecular weight and isoelectric point were 78798.75 Da and 8.64,respectively.The homology of flbA gene sequence between A.clavatus and A.nidulans was 69.9%,and the homology of amino acid sequence was 76.6%.The flbA gene knockout box and vector were constructed and 258 transformants of A.clavatus were obtained.A flbA gene deletion mutant was\{identified\}.The study would provide materials and lay a foundation for further study on the function of flbA gene.

关 键 词：棒曲霉 分生孢子 flbA基因 克隆 敲除载体 缺失突变株 

分 类 号：Q936[生物学—微生物学]