A highly efficient targeted recombination system for engineering linear chromosomes of industrial bacteria Streptomyces

The Journal of General and Applied Microbiology

Carton W. Chen¹  Hung-Yin Pan²  Chih-Hung Huang³ 

[1] Department of Life Sciences and Institute of Genome Sciences, National Yang-Ming University;Graduate Institution of Engineering Technology, National Taipei University of Technology;Institute of Biochemical and Biomedical Engineering, National Taipei University of Technology

关键词: genome engineering;    linear chromosome;    linear plasmid;    Streptomyces;    targeted recombination;   

DOI  :  10.2323/jgam.2017.11.004

学科分类：微生物学和免疫学

来源: Applied Microbiology, Molecular and Cellulrar Biosciences Research Foundation

【 摘 要 】

Soil bacteria Streptomyces are the most important producers of secondary metabolites, including most known antibiotics. These bacteria and their close relatives are unique in possessing linear chromosomes, which typically harbor 20 to 30 biosynthetic gene clusters of tens to hundreds of kb in length. Many Streptomyces chromosomes are accompanied by linear plasmids with sizes ranging from several to several hundred kb. The large linear plasmids also often contain biosynthetic gene clusters. We have developed a targeted recombination procedure for arm exchanges between a linear plasmid and a linear chromosome. A chromosomal segment inserted in an artificially constructed plasmid allows homologous recombination between the two replicons at the homology. Depending on the design, the recombination may result in two recombinant replicons or a single recombinant chromosome with the loss of the recombinant plasmid that lacks a replication origin. The efficiency of such targeted recombination ranges from 9 to 83% depending on the locations of the homology (and thus the size of the chromosomal arm exchanged), essentially eliminating the necessity of selection. The targeted recombination is useful for the efficient engineering of the Streptomyces genome for large-scale deletion, addition, and shuffling.

【 授权许可】

Unknown