Single-Homology-Arm Linear DNA Recombination by the Nonhomologous End Joining Pathway as a Novel and Simple Gene Inactivation Method: a Proof-of-Concept Study in Dietzia sp. Strain DQ12-45-1b

Author:

Lu Shelian1,Nie Yong1,Wang Meng1,Xu Hong-Xiu12,Ma Dong-Ling1,Liang Jie-Liang1,Wu Xiao-Lei1

Affiliation:

1. College of Engineering, Peking University, Beijing, People's Republic of China

2. College of Architecture and Environment, Sichuan University, Chengdu, People's Republic of China

Abstract

Many nonmodel Gram-positive bacteria lack efficient genetic manipulation systems, but they express genes encoding Ku and LigD. The NHEJ pathway in Dietzia sp. DQ12-45-1b was evaluated and was used to successfully knock out 11 genes in the genome. Since bioinformatic studies revealed that the putative genes encoding Ku and LigD ubiquitously exist in phylogenetically diverse bacteria and archaea, the single-homology-arm linear DNA recombination by the NHEJ pathway could be a potentially applicable genetic manipulation method for diverse nonmodel prokaryotic organisms.

Funder

National High Technology Research and Development Program

National High Technology Research and Development Program of China

National Natural Science Foundation of China

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

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