Dna purification kits

The media components used in the study were acquired from BTL (Łódź, Poland). Pierce™ Unstained Protein MW Marker, PageRuler™ Plus Prestained Protein Ladder and PageRuler™ Unstained Broad Range Protein Ladder were sourced from Thermo Fisher Scientific Baltics UAB (Vilnus, Lithuania). Marathon DNA polymerase and DNA purification kits were obtained from A&A Biotechnology (Gdynia, Poland). The restriction enzymes (REases) and Antarctic Phosphatase were purchased from New England Biolabs (Ipswich, MA, USA). T4 DNA Ligase was from Epicentre (Madison, WI, USA). The plasmid pGFPuv was purchased from Clontech/Takara Bio Inc. (Kusatsu, Shiga, Japan). E. coli DH5α cells [F⁻ φ80lacZ∆M15 ∆(lacZYA-argF) U169 deoR recA1 endA1 hsdR17 (r_K⁻, m_K⁺) phoA supE44 λ⁻thi-1 gyrA96 relA1 Mrr-] were used for clone selection (Life Technologies, Carlsbad, CA, USA). For protein expression, E. coli BL21(DE3) [F⁻ompT hsdSB(r_B⁻, m_B⁻) gal dcm (DE3)] from Life Technologies (Carlsbad, CA, USA) and the T7 promoter-based pET21d(+) vector from Novagen (Madison, WI, USA) were used. The other reagents and chemicals were sourced from POCh S.A. (Gliwice, Poland), Sigma-Aldrich (St. Louis, MO, USA), Fluka Chemie GmbH (Buchs, Switzerland) or AppliChem Inc. (St. Louis, MO, USA). The oligodeoxyribonucleotide synthesis and DNA sequencing were conducted at Genomed S.A. (Warsaw, Poland).

Oligonucleotides for site-directed mutagenesis and dNTPs were purchased from Oligo.pl (Warsaw, Poland) or Sigma Aldrich (Poland, Poznan). DNA sequencing was performed at the Laboratory for Nucleic Acid and Protein Detection (Pomeranian Science and Technology Park, Gdynia, Poland). Pfu polymerase was purchased from Fermentas. S-Sepharose, Superdex 75 PC 10/300, Superdex 75 PC 3.2/30 columns and Gel Filtration Low Molecular Weight Calibration Kits were from GE Healthcare (Warsaw, Poland). DNA purification kits were purchased from A&A Biotechnology (Gdansk, Poland). Unless otherwise specified, all reagents were of molecular biology or analytical grade.