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A flexible,modular and versatile functional part assembly toolkit for gene cluster engineering in Streptomyces

Synthetic and Systems Biotechnology 2024年第1期9卷 69-77页

作者： Xuejin Zhao Yeqing Zong Qiuli Lou Chenrui Qin Chunbo Lou CAS Key Laboratory of Pathogenic Microbiology and Immunology Institute of MicrobiologyChinese Academy of SciencesBeijing100101China CAS Key Laboratory of Microbial Physiological and Metabolic Engineering Institute of MicrobiologyChinese Academy of SciencesBeijing100101China Center for Cell and Gene Circuit Design Key Laboratory of Quantitative Synthetic BiologyShenzhen Institute of Synthetic BiologyShenzhen Institutes of Advanced TechnologyChinese Academy of Sciences1068 Xueyuan AvenueUniversity TownNanshanShenzhen518055China Peking-Tsinghua Joint Center for Life Sciences Peking UniversityBeijing100871China Center for Quantitative Biology Academy for Advanced Interdisciplinary StudiesSchool of PhysicsPeking UniversityBeijing100871China College of Life Sciences University of Chinese Academy of SciencesBeijing100149China

Streptomyces has enormous potential to produce novel natural products(NPs)as it harbors a huge reservoir of uncharacterized and silent natural product biosynthetic gene clusters(BGCs).However,the lack of efficient gene cluster engineering strategies has hampered the pace of new drug ***,we developed an easy-to-use,highly flexible DNA assembly toolkit for gene cluster *** DNA assembly toolkit is compatible with various DNA assembling approaches including Biobrick,Golden Gate,CATCH,yeast homologous recombination-based DNA assembly and homing endonuclease-mediated *** compatibility offers great flexibility in handling multiple genetic parts or refactoring large gene *** demonstrate the utility of this toolkit,we quantified a library of modular regulatory parts,and engineered a gene cluster(act)using characterized promoters that led to increased ***,this work provides a powerful part assembly toolkit that can be used for natural product discovery and optimization in Streptomyces.

关键词： Streptomyces standard toolkit DNA assembly Gene cluster Synthetic biology

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