title: Efficient and simple generation of multiple unmarked gene deletions in Mycobacterium smegmatis.
abstract: Research on mycobacterial genetics relies heavily on techniques for directed gene mutation, but genetic studies are often hampered by the difficulty of generating gene deletions in mycobacteria. We developed an efficient and improved deletion system, described here in detail, which can be used to construct multiple unmarked recombinants in mycobacteria. We tested this system by using it to sequentially delete four pairs of toxin-antitoxin genes in Mycobacterium smegmatis.
authors.affiliation: MOH Key Laboratory of Systems Biology of Pathogens, Institute of Pathogen Biology, and Center for Tuberculosis Research, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, P.R. China.
authors.forename: Xu-Jian
authors.initials: XJ
authors.lastname: Mao
authors.affiliation: MOH Key Laboratory of Systems Biology of Pathogens, Institute of Pathogen Biology, and Center for Tuberculosis Research, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, P.R. China.
authors.forename: Mei-Yi
authors.initials: MY
authors.lastname: Yan
authors.affiliation: MOH Key Laboratory of Systems Biology of Pathogens, Institute of Pathogen Biology, and Center for Tuberculosis Research, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, P.R. China.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhu
authors.affiliation: MOH Key Laboratory of Systems Biology of Pathogens, Institute of Pathogen Biology, and Center for Tuberculosis Research, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, P.R. China.
authors.forename: Xiao-Peng
authors.initials: XP
authors.lastname: Guo
authors.affiliation: MOH Key Laboratory of Systems Biology of Pathogens, Institute of Pathogen Biology, and Center for Tuberculosis Research, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, P.R. China.
authors.forename: Yi-Cheng
authors.initials: YC
authors.lastname: Sun
mesh_terms: D000992:Antitoxins / Q000235:genetics; D001427:Bacterial Toxins / Q000235:genetics; D017353:Gene Deletion; D055786:Gene Knockout Techniques / Q000379:methods*; D005798:Genes, Bacterial / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics; D008957:Models, Genetic; D009154:Mutation; D020102:Mycobacterium smegmatis / Q000235:genetics* / Q000254:growth & development; D010957:Plasmids / Q000235:genetics; D011995:Recombination, Genetic; D014169:Transformation, Bacterial
chemical_list: D000992:Antitoxins; D001427:Bacterial Toxins; D049452:Green Fluorescent Proteins