Pqe 30 ua vector

LukF-PV was amplified using LukF-recombinant primers (Table 1) and cloned into pQE-30 UA vector (Qiagen). Recombinant protein expression was induced according to the manufacturer's instructions and recombinant His-tagged LukF-PV was purified using a His-bind purification kit (Novagen).

Trypticase peptone yeast extract glucose (TPYG) and Luria Bertani (LB) medium were purchased from Difco Laboratories, USA. Escherichia coli SG13009, pQE30-UA vector, Ni–NTA resin, plasmid DNA purification kit (27,106), and PCR Kit were procured from Qiagen, USA. Pre-stained protein markers, unstained protein molecular weight marker, (SM0431), DNA ladder (SM0313) were obtained from Fermentas, USA. Primers were synthesized from Microsynth, Switzerland. Cell culture media components, monoclonal anti-polyhistidine antibody raised in mouse (H1029), HRP conjugate AP163P, Anti-His HRP antibody, mouse monoclonal antibody isotyping reagents (ISO2) and chicken anti-goat Ig antibody, 3, 3′-diaminobenzidine (DAB), and other required chemicals were purchased from Sigma Aldrich, USA. Cytokine ELISA kit & flowcytometry reagents were obtained from BD Biosciences, USA.