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Lomakin Y.A.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Zakharova M.Y.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Stepanov A.V.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Dronina M.A.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | And 21 more authors.
Molecular Immunology | Year: 2014

The mechanisms triggering most of autoimmune diseases are still obscure. Autoreactive B cells play a crucial role in the development of such pathologies and, in particular, production of autoantibodies of different specificities. The combination of deep-sequencing technology with functional studies of antibodies selected from highly representative immunoglobulin combinatorial libraries may provide unique information on specific features in the repertoires of autoreactive B cells. Here, we have analyzed cross-combinations of the variable regions of human immunoglobulins against the myelin basic protein (MBP) previously selected from a multiple sclerosis (MS)-related scFv phage-display library. On the other hand, we have performed deep sequencing of the sublibraries of scFvs against MBP, Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1), and myelin oligodendrocyte glycoprotein (MOG). Bioinformatics analysis of sequencing data and surface plasmon resonance (SPR) studies have shown that it is the variable fragments of antibody heavy chains that mainly determine both the affinity of antibodies to the parent autoantigen and their cross-reactivity. It is suggested that LMP1-cross-reactive anti-myelin autoantibodies contain heavy chains encoded by certain germline gene segments, which may be a hallmark of the EBV-specific B cell subpopulation involved in MS triggering. © 2014 The Authors. Source


Kurochkina V.B.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Sklyarenko A.V.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Satarova J.E.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Yarotsky S.V.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika
Bioprocess and Biosystems Engineering | Year: 2011

The article deals with experimental determination of ionization constants and solubility for the compounds (target products, initial β-lactams, acylating agents and by-products) involved in enzymatic synthesis of some therapeutically used aminopenicillins and aminocephalosporins, namely ampicillin, amoxicillin, cephalexin, cephadroxil, cephaloglycin, cefaclor, cefprozil, cefatrizine. Methodology of investigations and the evaluation of experimental data for the determination of ionization constants and solubility of the different type electrolytes are presented. Applications of the methods based on acid-base potentiometric titration and on determination of solubility-pH dependence of assayed substances are discussed. The original data on ionization constants and solubility of amoxicillin, cefprozil, cefatrizine, cephadroxil and initial β-lactams for production of cefaclor, cefprozil and cefatrizine, as well as solubility of by-product d-(-)-p-hydroxyphenylglycine are presented. Experimentally determined parameters and constants available in the literature for all abovementioned aminopenicillins and aminocephalosporins are collected. These data might be used for choice of the conditions of both processes: the enzymatic synthesis and the isolation of the product from reaction mixture. © 2011 Springer-Verlag. Source


Mordkovich N.N.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Manuvera V.A.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Veiko V.P.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Debabov V.G.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika
Applied Biochemistry and Microbiology | Year: 2012

A DNA fragment containing a promoter-operator and structural parts of the uridine phosphorylase gene from Shewanella oneidensis MR-1 was cloned. Cross-heterological expression of the udp genes from Sh. oneidensis MR-1 and Escherichia coli under the control of authentic regulatory regions is shown. The UDP protein accumulates in an active form in the cytoplasmic fraction of cells. The recombinant UDP protein from Sh. oneidensis MR-1 obtained by heterological expression was isolated and characterized. E. coli udp gene promoter activity was observed during heterological expression in Sh. oneidensis MR-1 cells under both aerobic and anaerobic conditions. © 2012 Pleiades Publishing, Ltd. Source


Khrul'nova S.A.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Maryshev I.V.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Kulikovsky A.D.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Manukhov I.V.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Zavilgelsky G.B.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika
Biologicheskie Membrany | Year: 2012

Structural features of lux operon of the psychrophilic marine luminescent bacteria Aliivibrio logei(strains KChl and BM1) isolated from the intestine of goby fish(White Sea and Sea of Okhotsk) are characterized. Bioluminescent characteristics of the strains are determined and compared with lux operon of the mesophilic marine luminescent bacteria A. fischeri. We conclude that in spite of differences in structure of lux operons of A. fischeri and A. logei they have similar sensitivities to autoinducer. Source


Yuzbashev T.V.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Larina A.S.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Vybornaya T.V.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Yuzbasheva E.Y.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | And 2 more authors.
Fungal Biology | Year: 2015

The vast number of repetitive genomic elements was identified in the genome of Rhizopus oryzae. Such genomic repeats can be used as homologous regions for integration of plasmids. Here, we evaluated the use of two different repeats: the short (575bp) rptZ, widely distributed (about 34 copies per genome) and the long (2053 bp) rptH, less prevalent (about 15 copies). The plasmid carrying rptZ integrated, but did so through a 2256-bp region of homology to the pyrG locus, a unique genomic sequence. Thus, the length of rptZ was below the minimal requirements for homologous strand exchange in this fungus. In contrast, rptH was used efficiently for homologous integration. The plasmid bearing this repeat integrated in multicopy fashion, with up to 25 copies arranged in tandem. The latter vector, pPyrG-H, could be a valuable tool for integration at homologous sequences, for such purposes as high-level expression of proteins. © 2015 The British Mycological Society. Source

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