title: High-Efficiency, Two-Step Scarless-Markerless Genome Genetic Modification in Salmonella enterica.
abstract: We present a two-step method for scarless-markerless genome genetic modification in Salmonella enterica based on the improved suicide plasmid pGMB152. The whole LacZYA gene can provide a lacZ-based blue/white screening strategy for fast selection of double-crossover mutants by allelic exchange. The high efficiency of this genetic engineering strategy permits the study of gene function by gene knockin, site-directed mutagenesis, and gene knockout to construct live attenuated vaccines.
authors.affiliation: Jiangsu Key Laboratory of Zoonosis, Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, 225009, China.
authors.forename: Shizhong
authors.initials: S
authors.lastname: Geng
authors.affiliation: Jiangsu Key Laboratory of Zoonosis, Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, 225009, China.
authors.forename: Qin
authors.initials: Q
authors.lastname: Tian
authors.affiliation: Jiangsu Key Laboratory of Zoonosis, Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, 225009, China.
authors.forename: Shuming
authors.initials: S
authors.lastname: An
authors.affiliation: Jiangsu Key Laboratory of Zoonosis, Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, 225009, China.
authors.forename: Zhiming
authors.initials: Z
authors.lastname: Pan
authors.affiliation: Jiangsu Key Laboratory of Zoonosis, Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, 225009, China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Chen
authors.affiliation: Jiangsu Key Laboratory of Zoonosis, Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, 225009, China. jiao@yzu.edu.cn.
authors.forename: Xinan
authors.initials: X
authors.lastname: Jiao
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D005818:Genetic Engineering / Q000379:methods*; D016680:Genome, Bacterial; D016254:Mutagenesis, Insertional; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D019779:Salmonella enterica / Q000235:genetics* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins