Expression, production, and characterization of full-length vitronectin in Escherichia coli

摘要: Vitronectin (VN) is one of the primary adhesive proteins in serum and serves to promote attachment spreading a wide variety cell types tissue culture plastic. In this study, pGEX2t expression vector was used express full-length human VN as GST-tagged fusion protein Escherichia coli. GST/VN production induced with IPTG found localize inclusion bodies. The bodies were isolated from lysates, washed once 2 M urea Triton X-100, then solubilized 8 presence reducing compound. Solubilized purified by heparin affinity chromatography refolded dialysis against phosphate buffered saline. Approximately 40 mg recovered 1L bacterial culture. Purified migrated at predicted molecular mass on SDS-PAGE recognized both anti-GST anti-VN antibodies. bound promoted adhesion, spreading, growth similar extent that observed plasma-derived VN. As such, recombinant bacteria represents rapid convenient method produce large quantities for cellular studies.