Song Ha-Yeon, Choi Dahye, Han Dong-Min, Kim Dae-Hyuk, Kim Jung-Mi
Department of Bio-Environmental Chemistry, Institute of Life Science and Natural Resources, Wonkwang University, Iksan, Chonbuk, Korea.
Division of Biological Sciences, Wonkwang University, Iksan, Chonbuk, Korea.
Mycobiology. 2018 Dec 17;46(4):429-439. doi: 10.1080/12298093.2018.1548806. eCollection 2018.
To develop a convenient promoter analysis system for fungi, a null-pigment mutant (NPG) of was used with the 4'-phosphopantetheinyl transferase (PPTase) gene, , which restores the normal pigmentation in , as a new reporter gene. The functional organization of serially deleted promoter regions of the gene and the gene in filamentous fungi was representatively investigated to establish a novel fungal promoter assay system that depends on color complementation of the NPG mutant with the PPTase gene. Several promoter regions of the and genes were fused to the gene containing the 1,034-bp open reading frame and the 966-bp 3' downstream region from the TAA, and the constructed fusions were introduced into the NPG mutant in to evaluate color recovery due to the transcriptional activity of the sequence elements. Serial deletion of the and promoter regions in this PPTase reporter assay system reaffirmed results in previous reports by using the fungal transformation step without a laborious verification process. This approach suggests a more rapid and convenient system than conventional analyses for fungal gene expression studies.
为开发一种适用于真菌的便捷启动子分析系统,使用了一种无色突变体(NPG),并将4'-磷酸泛酰巯基乙胺基转移酶(PPTase)基因作为新的报告基因,该基因可恢复其正常色素沉着。对丝状真菌中该基因和基因的串联缺失启动子区域的功能组织进行了代表性研究,以建立一种基于NPG突变体与PPTase基因颜色互补的新型真菌启动子检测系统。将该基因和基因的几个启动子区域与包含1034 bp开放阅读框和来自TAA的966 bp 3'下游区域的基因融合,并将构建的融合体导入中的NPG突变体中,以评估由于序列元件的转录活性导致的颜色恢复。在该PPTase报告基因检测系统中对该基因和启动子区域进行串联缺失,再次证实了先前报告中的结果,且无需繁琐的验证过程即可完成真菌转化步骤。该方法表明,与传统的真菌基因表达研究分析方法相比,它是一种更快速、便捷的系统。