3 phenoxybenzoic acid

T4 DNA ligase and restriction enzyme SfiI were obtained from New England Biolabs, Inc. (Beverly, MA). Phusion High-Fidelity DNA Polymerase, bacterial protein extraction reagent (B-PER), HisPur Ni-NTA resin, NuPAGE Bis-Tris Gel (the precast polyacrylamide gels designed to give optimal separation for a wide range of molecular weight proteins during gel electrophoresis) and chemically competent cells of E. coli BL21 (DE3) pLysS were from Thermo Fisher Scientific (Rockford, IL). Standards (3-phenoxybenzoic acid, 1, and its analogs, 3-phenoxybeneyl aldehyde and 3-phenoxybenzyl alcohol; permethrin, 2; cypermethrin, 3; deltamethrin, 4; fenpropathrin, 5; and phenothrin, 6) (Figure 1)), isopropyl-β-D-thiogalactopyranoside (IPTG), and p-nitrophenyl phosphate (pNPP) substrate were from Sigma (St. Louis, MO). The AttoPhos AP fluorescent substrate system was purchased from Roche (Pleasanton, CA). The vector pecan 45 containing AP gene was a generous gift from Dr. Jinny L. Liu and Dr. Ellen R. Goldman (Naval Research Laboratory, Center for Bio/Molecular Science and Engineering, Washington, DC). The AP-conjugated goat anti-mouse lgG was purchased from Jackson ImmunoResearch Inc. (West Grove, PA).