title: Construction of an engineered strain capable of degrading two isomeric nitrophenols via a sacB- and gfp-based markerless integration system.
abstract: In this study, a gfp-based novel markerless allelic exchange integration system was developed. By employing gfp gene and sacB gene as counter-selectable markers, an ortho-nitrophenol degradation operon (onpABC gene cluster) was successfully inserted into the chromosome of meta-nitrophenol utilizer Cupriavidus necator JMP134. Through two rounds of recombination, the engineered strain (strain JMP134-ONP) was directly selected from the plate by fluorescence screening and has the ability to degrade both ortho-nitrophenol and meta-nitrophenol, simultaneously. This relatively simple and efficient method can be used as an alternative strategy of allelic exchange insertion for the application of metabolic engineering in various bacterial strains, complementary to existing gene knock-in procedures.
authors.affiliation: Key Laboratory of Agricultural and Environmental Microbiology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Hu
authors.forename: Xuan
authors.initials: X
authors.lastname: Jiang
authors.forename: Jun-Jie
authors.initials: JJ
authors.lastname: Zhang
authors.forename: Ning-Yi
authors.initials: NY
authors.lastname: Zhou
mesh_terms: D001711:Biotransformation; D020118:Cupriavidus necator / Q000235:genetics* / Q000378:metabolism*; D005827:Genetics, Microbial / Q000379:methods; D060847:Metabolic Engineering / Q000379:methods*; D008967:Molecular Biology / Q000379:methods; D009596:Nitrophenols / Q000378:metabolism*; D012641:Selection, Genetic
chemical_list: D009596:Nitrophenols