title: [Influence of physiological state of Escherichia coli cells on the expression of soluble protein--recombinant analog of glycoprotein G of Herpes simplex virus of type 2].
abstract: It is shown that the recombinant protein GST-HSV2gG, containing the immunodominant regions of glycoprotein G of HSV-2 is accumulated in the form of inclusion bodies or in soluble form in the Escherichia coli BL21 (DE3) cells. The ratio between protein fractions varied depending on the physiological state of cells before biosynthesis. The kinetic parameters of bacterial populations were determined by mathematical modeling of growth curves based on the Verhulst logistic function. It was established that the induction of biosynthesis in the growth acceleration phase (at OD600 = 0.3) with 0.1 mM IPTG gives the maximum yield of soluble protein (26.75 mg/l or 17.6 mg/g biomass). The target protein was purified using the immobilized metal ion affinity and affinity chromatography technologies. Antigenic activity of the soluble form of recombinant protein GST-HSV2gG, was significantly (three times) higher than that of the protein purified from inclusion bodies (p < 0.05) and was comparable with the activity of the commercial analog (p > 0.05), that allows using this product in the immunosorbent test kits for diagnosis of IgG to HSV-2.
authors.forename: L N
authors.initials: LN
authors.lastname: Korshun
authors.forename: L N
authors.initials: LN
authors.lastname: Moĭsa
authors.forename: L A
authors.initials: LA
authors.lastname: Ganova
authors.forename: M I
authors.initials: MI
authors.lastname: Vudmaska
authors.forename: G V
authors.initials: GV
authors.lastname: Kovtoniuk
authors.forename: E K
authors.initials: EK
authors.lastname: Kiseleva
authors.forename: N Ia
authors.initials: NIa
authors.lastname: Spivak
mesh_terms: D000941:Antigens / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D008954:Models, Biological; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D012995:Solubility; D014759:Viral Envelope Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification
chemical_list: D000941:Antigens; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C045464:glycoprotein G, herpes simplex virus type 2; D007544:Isopropyl Thiogalactoside