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用于……的嵌合开放阅读框融合表型报告基因

A Chimeric ORF Fusion Phenotypic Reporter for .

作者信息

Phillips-Rose Louis S, Yu Chendi K, West Nicholas P, Fraser James A

机构信息

Australian Infectious Diseases Research Centre, School of Chemistry & Molecular Biosciences, The University of Queensland, Brisbane, QLD 4072, Australia.

出版信息

J Fungi (Basel). 2024 Aug 12;10(8):567. doi: 10.3390/jof10080567.

Abstract

The plethora of genome sequences produced in the postgenomic age has not resolved many of our most pressing biological questions. Correlating gene expression with an interrogatable and easily observable characteristic such as the surrogate phenotype conferred by a reporter gene is a valuable approach to gaining insight into gene function. Many reporters including , and the fluorescent proteins mRuby3 and mNeonGreen have been used across all manners of organisms. Described here is an investigation into the creation of a robust, synthetic, fusion reporter system for that combines some of the most useful fluorophores available in this system with the versatility of the counter-selectable nature of . The reporters generated include multiple composition and orientation variants, all of which were investigated for differences in expression. Evaluation of known promoters from the and genes was undertaken, elucidating novel expression tendencies of these biologically relevant regulators of transcription. Smaller than but providing multiple useful surrogate phenotypes for interrogation, the fusion ORF serves as a superior whole-cell assay compared to traditional systems. Ultimately, the work described here bolsters the array of relevant genetic tools that may be employed in furthering manipulation and understanding of the WHO fungal priority group pathogen .

摘要

后基因组时代产生的大量基因组序列并未解决我们许多最紧迫的生物学问题。将基因表达与可询问且易于观察的特征(如报告基因赋予的替代表型)相关联,是深入了解基因功能的一种有价值的方法。包括……以及荧光蛋白mRuby3和mNeonGreen在内的许多报告基因已在各种生物体中使用。本文描述了一项针对……创建强大的合成融合报告系统的研究,该系统将该系统中一些最有用的荧光团与……的反选择性质的多功能性结合起来。产生的报告基因包括多种组成和方向变体,所有这些都对表达差异进行了研究。对来自……和……基因的已知启动子进行了评估,阐明了这些生物学相关的转录调节因子的新表达趋势。融合开放阅读框比……小,但提供了多种有用的替代表型以供询问,与传统系统相比,它是一种更优越的全细胞分析方法。最终,本文所述的工作加强了一系列相关的遗传工具,这些工具可用于进一步操纵和了解世界卫生组织真菌重点病原体……

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e696/11355724/c96f1ec5a33a/jof-10-00567-g001.jpg

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