title: Rapid method of luxS and pfs gene inactivation in enterotoxigenic Escherichia coli and the effect on biofilm formation.
abstract: Rapid and efficient inactivation of a target gene in Escherichia coli chromosomes is required to investigate metabolic engineering. In the present study, a multiple gene inactivation approach was demonstrated in four strains of enterotoxigenic E. coli (ETEC), which are the predominant pathogenic bacteria causing piglet diarrhea, mediated by λ Red and Xer recombination. The chromosomal genes, luxS and pfs were inactivated using the multiple gene inactivation approach in the wild‑type strains of E. coli, K88, K99, 987P and F41. This indicated that dif sites may be reused to inactivate multiple chromosomal genes when no antibiotic‑resistant selectable markers remain. Following inactivation of luxS and pfs, the ability of ETEC to produce the quorum sensing signal, and induce auto‑inducer 2 activity and biofilm formation were significantly reduced. Furthermore, the multiple gene inactivation approach also exhibits a high recombination efficiency and follows a simple process.
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, Jilin 130122, P.R. China.
authors.forename: Xiaoxu
authors.initials: X
authors.lastname: Wang
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, Jilin 130122, P.R. China.
authors.forename: Shiyu
authors.initials: S
authors.lastname: Li
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, Jilin 130122, P.R. China.
authors.forename: Xiaoran
authors.initials: X
authors.lastname: Lu
authors.affiliation: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun, Jilin 130062, P.R. China.
authors.forename: Pan
authors.initials: P
authors.lastname: Hu
authors.affiliation: Changchun University of Science and Technology, Changchun, Jilin 130022, P.R. China.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Chen
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, Jilin 130122, P.R. China.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Li
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, Jilin 130122, P.R. China.
authors.forename: Zhaoyang
authors.initials: Z
authors.lastname: Bu
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, Jilin 130122, P.R. China.
authors.forename: Xulong
authors.initials: X
authors.lastname: Lang
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary Medicine, Academy of Military Medical Sciences, Changchun, Jilin 130122, P.R. China.
authors.forename: Xinglong
authors.initials: X
authors.lastname: Wang
mesh_terms: D018441:Biofilms; D002876:Chromosomes, Bacterial / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D054307:Enterotoxigenic Escherichia coli / Q000235:genetics*; D020868:Gene Silencing; D005798:Genes, Bacterial; D005819:Genetic Markers; D005821:Genetic Techniques; D005822:Genetic Vectors / Q000378:metabolism; D006714:Homoserine / Q000031:analogs & derivatives / Q000378:metabolism; D007783:Lactones / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic / Q000235:genetics; D017422:Sequence Analysis, DNA
chemical_list: D005819:Genetic Markers; D007783:Lactones; C091088:N-octanoylhomoserine lactone; D006714:Homoserine