Evrogen JSC

Moscow, Russia

Evrogen JSC

Moscow, Russia
SEARCH FILTERS
Time filter
Source Type

Bogdanov E.A.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Shagina I.,Evrogen JSC | Barsova E.V.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Kelmanson I.,Evrogen JSC | And 2 more authors.
Current Protocols in Molecular Biology | Year: 2010

The characterization of rare messages in cDNA libraries is complicated by the substantial variations that exist in the abundance levels of different transcripts in cells and tissues. The equalization (normalization) of cDNA is a helpful approach for decreasing the prevalence of abundant transcripts, thereby facilitating the assessment of rare transcripts. This unit provides a method for duplex-specific nuclease (DSN)based normalization, which allows for the fast and reliable equalization of cDNA, thereby facilitating the generation of normalized, full-length-enriched cDNA libraries, and enabling efficient RNA analyses. Curr. Protoc. Mol. Biol. 90:5.12.1-5.12.27. © 2010 John Wiley & Sons, Inc.


Goptar I.A.,Moscow State University | Shagin D.A.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Shagina I.A.,Evrogen JSC | Mudrik E.S.,Evrogen JSC | And 7 more authors.
Insect Biochemistry and Molecular Biology | Year: 2013

Prolyl carboxypeptidase (PRCP) is a lysosomal proline specific serine peptidase that also plays a vital role in the regulation of physiological processes in mammals. In this report, we isolate and characterize the first PRCP in an insect. PRCP was purified from the anterior midgut of larvae of a stored product pest, Tenebrio molitor, using a three-step chromatography strategy, and it was determined that the purified enzyme was a dimer. The cDNA of PRCP was cloned and sequenced, and the predicted protein was identical to the proteomic sequences of the purified enzyme. The substrate specificity and kinetic parameters of the enzyme were determined. The T. molitor PRCP participates in the hydrolysis of the insect's major dietary proteins, gliadins, and is the first PRCP to be ascribed a digestive function. Our collective data suggest that the evolutionary enrichment of the digestive peptidase complex in insects with an area of acidic to neutral pH in the midgut is a result of the incorporation of lysosomal peptidases, including PRCP. © 2013.


Shcherbo D.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Shemiakina I.I.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Ryabova A.V.,RAS A.M. Prokhorov General Physics Institute | Luker K.E.,University of Michigan | And 11 more authors.
Nature Methods | Year: 2010

Fluorescent proteins with emission wavelengths in the near-infrared and infrared range are in high demand for whole-body imaging techniques. Here we report near-infrared dimeric fluorescent proteins eqFP650 and eqFP670. To our knowledge, eqFP650 is the brightest fluorescent protein with emission maximum above 635 nm, and eqFP670 displays the most red-shifted emission maximum and high photostability. © 2010 Nature America, Inc. All rights reserved.


Shemiakina I.I.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Ermakova G.V.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Cranfill P.J.,Florida State University | Baird M.A.,Florida State University | And 13 more authors.
Nature Communications | Year: 2012

Multicolour labelling with fluorescent proteins is frequently used to differentially highlight specific structures in living systems. Labelling with fusion proteins is particularly demanding and is still problematic with the currently available palette of fluorescent proteins that emit in the red range due to unsuitable subcellular localization, protein-induced toxicity and low levels of labelling efficiency. Here we report a new monomeric red fluorescent protein, called FusionRed, which demonstrates both high efficiency in fusions and low toxicity in living cells and tissues. © 2012 Macmillan Publishers Limited. All rights reserved.


Mamedov I.Z.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Shagina I.A.,Evrogen JSC | Kurnikova M.A.,Evrogen JSC | Novozhilov S.N.,Evrogen JSC | And 2 more authors.
European Journal of Human Genetics | Year: 2010

A number of genetic systems for human genetic identification based on short tandem repeats or single nucleotide polymorphisms are widely used for crime detection, kinship studies and in analysis of victims of mass disasters. Here, we have developed a new set of 32 molecular genetic markers for human genetic identification based on polymorphic retroelement insertions. Allele frequencies were determined in a group of 90 unrelated individuals from four genetically distant populations of the Russian Federation. The mean match probability and probability of paternal exclusion, calculated based on population data, were 5.53 × 10 14 and 99.784%, respectively. The developed system is cheap and easy to use as compared to all previously published methods. The application of fluorescence-based methods for allele discrimination allows to use the human genetic identification set in automatic and high-throughput formats. © 2010 Macmillan Publishers Limited All rights reserved.


Serebrovskaya E.O.,Nizhny Novgorod State Medical Academy | Gorodnicheva T.V.,Evrogen JSC | Ermakova G.V.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Solovieva E.A.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | And 6 more authors.
Biochemical Journal | Year: 2011

Proteins of the GFP (green fluorescent protein) family are widely used as passive reporters for live cell imaging. In the present study we used H2B (histone H2B)-tKR (tandem KillerRed) as an active tool to affect cell division with light. We demonstrated that H2B-tKR-expressing cells behave normally in the dark, but transiently cease proliferation following green-light illumination. Complete light-induced blockage of cell division for approx. 24 h was observed in cultured mammalian cells that were either transiently or stably transfected with H2B-tKR. Illuminated cells then returned to normal division rate. XRCC1 (X-ray cross complementing factor 1) showed immediate redistribution in the illuminated nuclei of H2B-tKR-expressing cells, indicating massive light-induced damage of genomic DNA. Notably, nondisjunction of chromosomes was observed for cells that were illuminated during metaphase. In transgenic Xenopus embryos expressing H2B-tKR under the control of tissue-specific promoters, we observed clear retardation of the development of these tissues in green-light-illuminated tadpoles. We believe that H2B-tKR represents a novel optogenetic tool, which can be used to study mitosis and meiosis progression per se, as well as to investigate the roles of specific cell populations in development, regeneration and carcinogenesis in vivo. © The Authors Journal compilation © 2011 Biochemical Society.


Shagina I.,Evrogen JSC | Bogdanova E.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Mamedov I.Z.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | Lebedev Y.,RAS Shemyakin Ovchinnikov Institute of Bioorganic Chemistry | And 2 more authors.
BioTechniques | Year: 2010

An application of duplex-specific nuclease (DSN) normalization technology to whole-genome shotgun sequencing of genomes with a large proportion of repetitive DNA is described. The method uses a thermostable DSN from the Kamchatka crab that specifically hydrolyzes dsDNA. In model experiments on human genomic DNA, we demonstrated that DSN normalization of double-stranded DNA formed during C0t analysis is effective against abundant repetitive sequences with high sequence identity, while retaining highly divergent repeats and coding regions at baseline levels. Thus, DSN normalization applied to C 0t analysis can be used to eliminate evolutionarily young repetitive elements from genomic DNA before sequencing, and should prove invaluable in studies of large eukaryotic genomes, such as those of higher plants.


Savitsky A.P.,RAS A.N. Bach Institute of Biochemistry | Rusanov A.L.,RAS A.N. Bach Institute of Biochemistry | Zherdeva V.V.,RAS A.N. Bach Institute of Biochemistry | Gorodnicheva T.V.,Evrogen JSC | And 2 more authors.
Theranostics | Year: 2012

We report a new technique to detect enzyme activity inside cells. The method based on Fluorescence Lifetime Imaging (FLIM) technology allows one to follow sensor cleavage by proteolytic enzyme caspase-3. Specifically, we use the FLIM FRET of living cells via the confocal fluorescence microscopy. A specially designed lentivector pLVT with the DNA fragment of TagRFP-23-KFP was applied for transduction of A549 cell lines. Computer simulations are carried out to estimate FRET efficiency and to analyze possible steric restrictions of the re-action between the substrate TagRFP-23-KFP and caspase-3 dimer. Successful use of the fuse protein TagRFP-23-KFP to register the caspase-3 activation based on average life-time measurements is demonstrated. We show that the average life-time distribution is dramat-ically changed for cells with the modified morphology that is typical for apoptosis. Namely, the short-lived component at 1.8-2.1 ns completely disappears and the long-lived component appears at 2.4-2.6 ns. The latter is a fingerprint of the TagRFP molecule released after cleavage of the TagRFP-23-KFP complex by caspase-3. Analysis of life-time distributions for population of cells allows us to discriminate apoptotic and surviving cells within single frame and to peform statistical analysis of drug efficiency. This system can be adjusted for HTS by using special readers oriented on measurements of fluorescence life-time. © Ivyspring International Publisher.


Patent
Evrogen Jsc | Date: 2010-05-10

Nucleic acid molecules encoding improved fluorescent mutants of the mKate2 protein, variants and derivatives thereof are provided, as well as proteins and peptides encoded by these nucleic acids. Also provided are proteins that are substantially similar to, or derivatives, homologues, or mutants of, the above-referenced specific proteins. Also provided are fragments of the nucleic acids and the peptides encoded thereby, as well as antibodies specific to the proteins and peptides of the invention. In addition, host-cells, stable cell lines and transgenic organisms comprising above-referenced nucleic acid molecules are provided. The subject protein and nucleic acid compositions find use in a variety of different applications and methods, particularly for labeling of biomolecules, cells or cell organelles. Finally, kits for use in such methods and applications are provided.


Patent
Evrogen Jsc | Date: 2013-06-06

Nucleic acid molecules encoding improved fluorescent mutants of the mKate2 protein, variants and derivatives thereof are provided, as well as proteins and peptides encoded by these nucleic acids. Also provided are proteins that are substantially similar to, or derivatives, homologues, or mutants of, the above-referenced specific proteins. Also provided are fragments of the nucleic acids and the peptides encoded thereby, as well as antibodies specific to the proteins and peptides of the invention. In addition, host-cells, stable cell lines and transgenic organisms comprising above-referenced nucleic acid molecules are provided. The subject protein and nucleic acid compositions find use in a variety of different applications and methods, particularly for labeling of biomolecules, cells or cell organelles. Finally, kits for use in such methods and applications are provided.

Loading Evrogen JSC collaborators
Loading Evrogen JSC collaborators