ABSTRACT
Production of recombinant human apolipoprotein A-I (apoA-I) in E. coli cells is described and its biological properties are compared with those of natural protein. Recombinant apoA-I was isolated as a chimeric polypeptide and then processed to a mature form apoA-I (rapo-I). We studied the ability of the resulting protein to penetrate into hepatocyte nuclei and regulate the rate of DNA biosynthesis in complex with estriol. Penetration of rapoA-I conjugated with FITC into hepatocyte nuclei was demonstrated. rapoA-I-estriol and apoA-I-estriol complexes induced similar increase in DNA biosynthesis rate in isolated hepatocytes, which confi rms functional similarity of the obtained recombinant mature protein (rapoA-I) and native human apoA-I.
Subject(s)
Apolipoprotein A-I/pharmacology , Apolipoprotein A-I/genetics , Apolipoprotein A-I/isolation & purification , Cell Nucleolus/metabolism , Cell Nucleus/metabolism , Cells, Cultured , DNA Replication/drug effects , Escherichia coli , Estriol/pharmacology , Fluorescein-5-isothiocyanate , Fluorescent Dyes , Hepatocytes/drug effects , Hepatocytes/metabolism , Humans , Recombinant Fusion Proteins/isolation & purification , Recombinant Fusion Proteins/pharmacologyABSTRACT
The structural proteins OspC, FlaB, FlaA and DbpB of agent of ixodic tick borreliosis are one of main antigens inducing humoral immunity at initial stages of disease. Owing to it, the task was stated to evaluate antigen activity of recombinant proteins OspC (OscP-Bg), fragment of FlaB (f-FlaB) and DbpB of genospecies B. garinii and OspC (OscC-Ba) of genospecies of B. afzelii of West Siberian isolates of Borrelia Burgdorferi S.L. for their possible application as antigens for serological diagnostic of ixodic tick borreliosis The recombinant proteins OscP-Bg, OscC-Ba, f-FlaB, FlaA and DbpB are analyzed using technique of enzymoimmunoassay to detect ability to bound antibodies of serums of patients with ixodic tick borreliosis with localized and disseminated stage of infection. The difference of their sensitivity as antigens during detection of specific antibodies in blood serum of patients with ixodic tick borreliosis was demonstrated In serum of patients with ixodic tick borreliosis with disseminated stage of infection the level of specific IgM and IgG antibodies reacting with OscP-Bg, OscC-Ba, f-FlaB, FlaA and DbpB is within the limits 15.7-52.6% for IgG. The results of enzymoimmunoassay applied to patients with ixodic tick borreliosis for detection of IgM and IgG in serum demonstrated that OscP-Bg and f-FlaB determined the highest antigen activity with antigens. The study results make it possible to consider these proteins as perspective components for development of immune enzyme test system of diagnostic of ixodic tick borreliosis.
Subject(s)
Bacterial Proteins/immunology , Lyme Disease/diagnosis , Antibodies, Bacterial/immunology , Borrelia burgdorferi/immunology , Borrelia burgdorferi/isolation & purification , Humans , Immunoglobulin G/immunology , Immunoglobulin M/immunology , Lyme Disease/epidemiology , Lyme Disease/immunology , Recombinant Proteins/immunology , Serologic Tests , SiberiaABSTRACT
A sensitive biosensor test system was developed for evaluation of premutation effects of propellants on the cell genome.
