title: Bxb1 integrase serves as a highly efficient DNA recombinase in rapid metabolite pathway assembly.
abstract: Phage-encoded serine integrases are widely used in genetic engineering. They also have the potential to serve as efficient DNA assemblers, demonstrated by the method of site-specific recombination-based tandem assembly (SSRTA) that can combine biological parts into devices, pathways, and systems. Here, four serine integrases, ϕBT1, TG1, ϕRv1, and Bxb1, were investigated to ascertain their in vitro DNA assembly activities. Bxb1 integrase displayed the highest efficiency to obtain final products. Thus, we conclude that Bxb1 integrase is an excellent choice for DNA assembly in vitro Using this enzyme and its recognition sites, BioBrick standards were designed that are compatible with the SSRTA method for module addition. A rapid and efficient procedure was developed for the assembly of a multigene metabolic pathway in one step, directly from non-cutting plasmids containing the gene fragments. This technique is easy and convenient, and would be of interest to the synthetic biology community.
authors.affiliation: Collaborative Innovation Center for Genetics and Development, State Key Laboratory of Genetic Engineering, Department of Microbiology, School of Life Sciences, Fudan University, Shanghai 200438, China.
authors.forename: Xianwei
authors.initials: X
authors.lastname: Wang
authors.affiliation: Collaborative Innovation Center for Genetics and Development, State Key Laboratory of Genetic Engineering, Department of Microbiology, School of Life Sciences, Fudan University, Shanghai 200438, China.|CAS Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.|Institute of Quality and Standard for Agro-Products, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
authors.forename: Biao
authors.initials: B
authors.lastname: Tang
authors.affiliation: Department of Microbiology and Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, New Territories, Hong Kong SAR, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Ye
authors.affiliation: Collaborative Innovation Center for Genetics and Development, State Key Laboratory of Genetic Engineering, Department of Microbiology, School of Life Sciences, Fudan University, Shanghai 200438, China.
authors.forename: Yayi
authors.initials: Y
authors.lastname: Mao
authors.affiliation: Collaborative Innovation Center for Genetics and Development, State Key Laboratory of Genetic Engineering, Department of Microbiology, School of Life Sciences, Fudan University, Shanghai 200438, China.
authors.forename: Xiaolai
authors.initials: X
authors.lastname: Lei
authors.affiliation: Collaborative Innovation Center for Genetics and Development, State Key Laboratory of Genetic Engineering, Department of Microbiology, School of Life Sciences, Fudan University, Shanghai 200438, China gpzhao@sibs.ac.cn xmding74@fudan.edu.cn.|CAS Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.|Department of Microbiology and Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, New Territories, Hong Kong SAR, China.
authors.forename: Guoping
authors.initials: G
authors.lastname: Zhao
authors.affiliation: Collaborative Innovation Center for Genetics and Development, State Key Laboratory of Genetic Engineering, Department of Microbiology, School of Life Sciences, Fudan University, Shanghai 200438, China gpzhao@sibs.ac.cn xmding74@fudan.edu.cn.
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Ding
mesh_terms: D004926:Escherichia coli / Q000235:genetics*; D019426:Integrases / Q000378:metabolism*; D010957:Plasmids; D011994:Recombinant Proteins / Q000235:genetics; D045522:Recombinases / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D045522:Recombinases; D019426:Integrases
keywords: BioBrick; Bxb1 serine integrase; DNA assembly; site-specific recombination