Primestar hs dna polymerase and restriction enzymes

Supplementary Table 1 lists the strains used in this study. Escherichia coli NST37 (ATCC31882) was lysogenized using λDE3 Lysogenization kits (Novagen, Madison, WI, USA) to generate NST37 (DE3). We obtained 4APhe from Sigma Aldrich (St. Louis, MO, USA) and 4ACA, 4APE and 4APEA from Tokyo Chemical Industry (Tokyo, Japan). Plasmids were constructed using PrimeSTAR HS DNA polymerase and restriction enzymes (Takara Bio Inc., Shiga, Japan).

Materials and kits were purchased from the following sources: TIANamp Blood DNA Midi Kit (TIANGEN, Beijing, China); PrimeSTAR HS DNA polymerase and restriction enzymes (Takara Bio, Inc.; Otsu, Shiga, Japan); Spodoptera frugiperda (Sf)21 insect cells, Sf-900TM III SFM insect culture medium and Bac to-Bac Baculovirus Expression System (Invitrogen, Carlsbad, CA, United States); Mouse monoclonal anti-OR antibody (Santa Cruz Biotechnology, Dallas, Texas, United States); Rabbit polyclonal anti-CYP2C9 antibody (Abcam, Cambridge, United Kingdom); Super Signal West Pico Trial Kit (Thermo Scientific, Rockford, IL, United States); 4-Hydroxytolbutamide, 4-hydroxydiclofenac, and telmisartan (Toronto Research Chemicals, Inc.; Toronto, Ontario, Canada); Diclofenac and chlorpropamide (Tokyo Chemical Industry Co., Ltd.; Tokyo, Japan); Tolbutamide, losartan and E-3174 (Sigma-Aldrich, St. Louis, MO, United States); NADPH-regenerating system (Promega, Madison, WI, United States); High-pressure liquid chromatography-grade solvents (Fisher Scientific Co.; Fair Lawn, NJ, United States). All other chemicals and reagents were of analytical grade or the highest commercially available quality.