title: Transformation and characterization of an arsenic gene operon from urease-positive thermophilic Campylobacter (UPTC) in Escherichia coli.
abstract: An arsenate susceptibility test was performed with transformed and cultured Escherichia coli DH5α cells, which carried recombinant DNA of full-length arsenic (ars) operon, namely a putative membrane permease, ArsP; a transcriptional repressor, ArsR; an arsenate reductase, ArsC; and an arsenical-resistance membrane transporter, Acr3, from the Japanese urease-positive thermophilic Campylobacter lari (UPTC) CF89-12. The E. coli DH5α transformant showed reduced susceptibility to arsenate (~1536 μg/mL), compared to the control. Thus, these ars four-genes from the UPTC CF89-12 strain cells could confer a reduced susceptibility to arsenate in the transformed and E. coli DH5α cells. E. coli transformants with truncated ars operons, acr3 (acr3) and arsC-acr3 (∆arsC-acr3), of the ars operon, showed an MIC value of 384 μg/mL (~384 μg/mL), similar to the E. coli cells which carried the pGEM-T vector (control). Reverse transcription PCR confirmed in vivo transcription of recombinant full-length ars operon and deletion variants (∆acr3 and ∆arsC-acr3) in the transformed E. coli cells.
authors.affiliation: Laboratory of Molecular Biology, Graduate School of Environmental Health Sciences, Azabu University, Sagamihara, 252-5201, Japan. matsuda@azabu-u.ac.jp.
authors.forename: M
authors.initials: M
authors.lastname: Matsuda
authors.affiliation: Laboratory of Molecular Biology, Graduate School of Environmental Health Sciences, Azabu University, Sagamihara, 252-5201, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Kuribayashi
authors.affiliation: Laboratory of Molecular Biology, Graduate School of Environmental Health Sciences, Azabu University, Sagamihara, 252-5201, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Yamamoto
authors.affiliation: Department of Bacteriology, Northern Ireland Public Health Laboratory, Belfast City Hospital, Belfast, BT9 7AD, Northern Ireland, UK.
authors.forename: B C
authors.initials: BC
authors.lastname: Millar
authors.affiliation: Department of Bacteriology, Northern Ireland Public Health Laboratory, Belfast City Hospital, Belfast, BT9 7AD, Northern Ireland, UK.|School of Biomedical Sciences, Ulster University, Co. Londonderry, Coleraine, BT52 1SA, Northern Ireland, UK.|School of Medicine, Dentistry and Biomedical Sciences, Queen's University, Belfast, BT7 1NN, Northern Ireland, UK.
authors.forename: J E
authors.initials: JE
authors.lastname: Moore
mesh_terms: D001151:Arsenic / Q000378:metabolism* / Q000633:toxicity; D044863:Campylobacter lari / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling; D053858:Metabolic Networks and Pathways / Q000235:genetics*; D008826:Microbial Sensitivity Tests; D009876:Operon; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014170:Transformation, Genetic
chemical_list: D001151:Arsenic