Engineering cell factories for producing biofuels and pharmaceuticals has spurred great interests to develop rapid and efficient synthetic biology tools customized for modular pathway engineering.Along the way,combina...Engineering cell factories for producing biofuels and pharmaceuticals has spurred great interests to develop rapid and efficient synthetic biology tools customized for modular pathway engineering.Along the way,combinatorial gene expression control through modification of regulatory element offered tremendous opportunity for fine-tuning gene expression and generating digital-like genetic circuits.In this report,we present an efficient evolutionary approach to build a range of regulatory control elements.The reported method allows for rapid construction of promoter,5'UTR,terminator and trans-activating RNA libraries.Synthetic overlapping oligos with high portion of degenerate nucleotides flanking the regulatory element could be efficiently assembled to a vector expressing fluorescence reporter.This approach combines high mutation rate of the synthetic DNA with the high assembly efficiency of Gibson Mix.Our constructed library demonstrates broad range of transcriptional or translational gene expression dynamics.Specifically,both the promoter library and 50UTR library exhibits gene expression dynamics spanning across three order of magnitude.The terminator library and trans-activating RNA library displays relatively narrowed gene expression pattern.The reported study provides a versatile toolbox for rapidly constructing a large family of prokaryotic regulatory elements.These libraries also facilitate the implementation of combinatorial pathway engineering principles and the engineering of more efficient microbial cell factory for various biomanufacturing applications.展开更多
肌萎缩侧索硬化症(amyotrophic lateral sclerosis,ALS)是目前无有效治疗的一种神经退行性疾病,其发病机制不清。近期研究发现反式激活应答DNA结合蛋白43(transactive response element DNA binding protein-43,TDP-43)在包括ALS、额颞...肌萎缩侧索硬化症(amyotrophic lateral sclerosis,ALS)是目前无有效治疗的一种神经退行性疾病,其发病机制不清。近期研究发现反式激活应答DNA结合蛋白43(transactive response element DNA binding protein-43,TDP-43)在包括ALS、额颞叶痴呆等神经退行性疾病中起重要作用。本文总结了近10年来对TDP-43结构的最新认识及其在神经退行性疾病发病机制的作用与潜在的治疗靶点,以期为中西医结合诊治ALS提供一定的基础研究依据。展开更多
基金The authors would like to acknowledge the Department of Chemical,Biochemical and Environmental Engineering,College of Engineering and Information Technology,Office of the Vice President for Research(gratn number 10145-1113-021-STRT7XUPMAIN)at University of Maryland Baltimore County for funding support.
文摘Engineering cell factories for producing biofuels and pharmaceuticals has spurred great interests to develop rapid and efficient synthetic biology tools customized for modular pathway engineering.Along the way,combinatorial gene expression control through modification of regulatory element offered tremendous opportunity for fine-tuning gene expression and generating digital-like genetic circuits.In this report,we present an efficient evolutionary approach to build a range of regulatory control elements.The reported method allows for rapid construction of promoter,5'UTR,terminator and trans-activating RNA libraries.Synthetic overlapping oligos with high portion of degenerate nucleotides flanking the regulatory element could be efficiently assembled to a vector expressing fluorescence reporter.This approach combines high mutation rate of the synthetic DNA with the high assembly efficiency of Gibson Mix.Our constructed library demonstrates broad range of transcriptional or translational gene expression dynamics.Specifically,both the promoter library and 50UTR library exhibits gene expression dynamics spanning across three order of magnitude.The terminator library and trans-activating RNA library displays relatively narrowed gene expression pattern.The reported study provides a versatile toolbox for rapidly constructing a large family of prokaryotic regulatory elements.These libraries also facilitate the implementation of combinatorial pathway engineering principles and the engineering of more efficient microbial cell factory for various biomanufacturing applications.
文摘肌萎缩侧索硬化症(amyotrophic lateral sclerosis,ALS)是目前无有效治疗的一种神经退行性疾病,其发病机制不清。近期研究发现反式激活应答DNA结合蛋白43(transactive response element DNA binding protein-43,TDP-43)在包括ALS、额颞叶痴呆等神经退行性疾病中起重要作用。本文总结了近10年来对TDP-43结构的最新认识及其在神经退行性疾病发病机制的作用与潜在的治疗靶点,以期为中西医结合诊治ALS提供一定的基础研究依据。
