Quantitative characterization of filamentous fungal promoters on a single-cell resolution to discover cryptic natural products  

作　　者：Peng-Lin Wei Jie Fan Jingwen Yu Zihui Ma Xian Guo Nancy PKeller Erwei Li Chunbo Lou Wen-Bing Yin 

机构地区：[1]State Key Laboratory of Mycology,Institute of Microbiology,Chinese Academy of Sciences,Beijing 100101,China [2]Savaid Medical School,University of Chinese Academy of Sciences,Beijing 100049,China [3]Department of Medical Microbiology and Immunology,University of Wisconsin-Madison,Madison,WI 53706,USA [4]CAS Key Laboratory of Quantitative Engineering Biology,Shenzhen Institute of Synthetic Biology,Shenzhen Institutes of Advanced Technology,Chinese Academy of Sciences,Shenzhen 518055,China

出　　处：《Science China(Life Sciences)》2023年第4期848-860,共13页中国科学（生命科学英文版）

基　　金：supported by the National Key Research and Development Program of China(2020YFA0907800);the National Natural Science Foundation of China(31861133004,81872771);the Key Research Program of Frontier Sciences,CAS(ZDBS-LY-SM016);the Biological Resources Programme,Chinese Academy of Sciences(KFJBRP-009-005);the China Postdoctoral Science Foundation(YJ20200201,2020M680720,2022T150689)。

摘　　要：Characterization of filamentous fungal regulatory elements remains challenging because of time-consuming transformation technologies and limited quantitative methods.Here we established a method for quantitative assessment of filamentous fungal promoters based on flow cytometry detection of the superfolder green fluorescent protein at single-cell resolution.Using this quantitative method,we acquired a library of 93 native promoter elements from Aspergillus nidulans in a high-throughput format.The strengths of identified promoters covered a 37-fold range by flow cytometry.P_(zipA) and P_(sltA)were identified as the strongest promoters,which were 2.9-and 1.5-fold higher than that of the commonly used constitutive promoter P_(gpdA).Thus,we applied P_(zipA)and P_(sltA)to activate the silent nonribosomal peptide synthetase gene Afpes1 from Aspergillus fumigatus in its native host and the heterologous host A.nidulans.The metabolic products of Afpes1 were identified as new cyclic tetrapeptide derivatives,namely,fumiganins A and B.Our method provides an innovative strategy for natural product discovery in fungi.

关 键 词：fungal promoter quantitative characterization single-cell resolution cryptic natural product activation of silent gene cluster 

分 类 号：Q933[生物学—微生物学]