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Immunogenic and protective activity of recombinant chimeric Toxoplasma gondii proteins
PublikacjaToxoplasmosis, one of the most common parasitic invasions worldwide, may pose a great threat for individuals with weakened immune system. The parasite also causes considerable economic losses due to infection of livestock. To date there is no efficient immunoprophylaxis for humans and animals, which would provide long-lasting protection against T. gondii invasion. Chimeric proteins, containing several selected fragments of parasite...
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The Immunogenic and Immunoprotective Activities of Recombinant Chimeric T. gondii Proteins Containing AMA1 Antigen Fragments
PublikacjaToxoplasmosis, one of the most common parasitoses worldwide, is potentially dangerous for individuals with a weakened immune system, but specific immunoprophylaxis intended for humans is still lacking. Thus, efforts have been made to create an efficient universal vaccine for both animals and humans to overcome the shortcomings of currently used treatment methods and protect all hosts against toxoplasmosis. The current work represents...
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Borrelia burgdorferi BmpA-BBK32 and BmpA-BBA64: New Recombinant Chimeric Proteins with Potential Diagnostic Value
PublikacjaCurrently, the diagnosis of Lyme disease is based mostly on two-tiered serologic testing. In the new generation of immunoenzymatic assays, antigens comprise whole-cell lysates of members of the Borrelia burgdorferi sensu lato (s.l.) species complex, with the addition of selected recombinant proteins. Due to the high diversity of members of the B. burgdorferi s.l. genospecies and the low degree of conservation among the amino acid...
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Toxoplasma gondii Recombinant antigen AMA1: Diagnostic Utility of Protein Fragments for the Detection of IgG and IgM Antibodies
PublikacjaToxoplasma gondii is an important zoonotic protozoan that infects a wide variety of vertebrates as intermediate hosts. For this reason, the diagnosis of this disease is very important and requires continuous improvement. One possibility is to use recombinant antigens in serological tests. Apical membrane antigen 1 (AMA1), a protein located in specific secretory organelles (micronemes) of T. gondii, is very interesting in regard...
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Hyperosmolarity adversely impacts recombinant protein synthesis by Yarrowia lipolytica—molecular background revealed by quantitative proteomics
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Novel monovalent and multivalent recombinant proteins of Borrelia burgdorferi sensu lato with potential diagnostic value – construction and biotechnological production
PublikacjaThe diverse antigenic structure of Borrelia burgdorferi sensu lato (s.l.) and the low degree of protein sequence conservation between genospecies causes many limitations in serodiagnosis of Lyme disease (LD). Using expression systems based on Escherichia coli, five monovalent B. burgdorferi s.l. recombinant proteins were produced. i.e., BB0108, BB0126, BB0298, BB0323, BB0689 (each in three variants derived from Borrelia afzelii,...
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PROTEIN EXPRESSION AND PURIFICATION
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Evaluation of long‑term immunity and protection against T. gondii after immunization with multivalent recombinant chimeric T. gondii proteins
PublikacjaToxoplasmosis caused by the opportunistic, cosmopolitan protozoan Toxoplasma gondii is one of the most common parasitoses in the world. Although it may prove dangerous or even fatal for immunocompromised individuals, immunoprophylaxis for humans is still nonexistent. Thus, the aim of the current work was to assess the ability of two immunogenic recombinant chimeric T. gondii proteins, SAG2-GRA1-ROP1 (SGR) and SAG1-MIC1-MAG1-GRA2...
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Function and Frustration of Multi-Drug ABC Exporter Protein and Design of Model Proteins for Drug Delivery Using Protein Hydration Thermodynamics
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PROTEINS-STRUCTURE FUNCTION AND BIOINFORMATICS
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Elastic-contractile model proteins: Physical chemistry, protein function and drug design and delivery
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How proteins bind to DNA: target discrimination and dynamic sequence search by the telomeric protein TRF1
PublikacjaTarget search as performed by DNA-binding proteins is a complex process, in which multiple factors contribute to both thermodynamic discrimination of the target sequence from overwhelmingly abundant off-target sites and kinetic acceleration of dynamic sequence interrogation. TRF1, the protein that binds to telomeric tandem repeats, faces an intriguing variant of the search problem where target sites are clustered within short fragments...
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Isolation and properties of recombinant trehalose synthase
PublikacjaBiosynthesis of trehalose is carried out via various matabolic pathways, including the use of trehalose synthase. Bacteria of the genus Deionococcus are microorganisms which produce trehalose synthase catalizing conversion of maltose into trehalose. In this work recombinant trehalose synthase from D.radiodurans was characterized.
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Probiotics and Antimicrobial Proteins
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Toxoplasma gondii: Enzyme-Linked Immunosorbent Assay using different fragments of recombinant microneme protein 1 (MIC1) for detection of immunoglobulin G antibodies
PublikacjaTrzy rekombinantowe fragmenty białka antygenowego MIC1 Toxoplasma gondii (r-MIC1ex2, r-MIC1ex34 i r-MIC1) zostały wyprodukowane jako białka fuzyjne (zawierające dwie domeny oligohistydynowe na N- i C-końcu) w komórkach bakteryjnych Escherichia coli. Homogenne preparaty antygenów rekombinantowych otrzymane z zastosowaniem jednoetapowego oczyszczania metodą chromatografii metalopowinowactwa, wykorzystano następnie do immunoidentyfikacji...
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Prime-Boost Vaccination with Toxoplasma Lysate Antigen, but Not with a Mixture of Recombinant Protein Antigens, Leads to Reduction of Brain Cyst Formation in BALB/c Mice
PublikacjaIntroduction Infection with the ubiquitous parasite Toxoplasma gondii is a threat for immunocompro- mised patients and pregnant women and effective immune-prophylaxis is still lacking. Methods Here we tested a mixture of recombinant T . gondii antigens expressed in different develop- mental stages, i.e., SAG1, MAG1 and GRA7 (SMG), and a lysate derived from T . gondii tachyzoites (TLA) for prophylactic vaccination against cyst formation....
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Extension of the Unres Package for Physics-Based Coarse-Grained Simulations of Proteins and Protein Complexes to Very Large Systems
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Proteins
PublikacjaStruktura białek wpływa na ich wartość odżywczą, właściwości funkcjonalne wżywności oraz reakcje w surowcach i produktach żywnościowych. Warunki prze-chowywania oraz obróbki w przemyśle i w czasie przygotowywania kulinarnegowywołują przemiany białek istotne dla jakości żywności. Przez celowe, enzy-matyczne i chemiczne modyfikowanie składu i struktury białek można wpływaćna ich właściwości funkcjonalne w żywności i wartość...
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The first study on the usefulness of recombinant tetravalent chimeric proteins containing fragments of SAG2, GRA1, ROP1 and AMA1 antigens in the detection of specific anti-Toxoplasma gondii antibodies in mouse and human sera
PublikacjaThis study presents an evaluation of four tetravalent recombinant chimeric proteins containing fragments of the Toxoplasma gondii antigens, SAG2, GRA1, ROP1 and AMA1, as potential replacements of a the soluble, whole-cell tachyzoite lysate (TLA) used in serological assays. Recombinant chimeric proteins (SAG2-GRA1-ROP1-AMA1N, AMA1N-SAG2-GRA1-ROP1, AMA1C-SAG2-GRA1-ROP1, and AMA1-SAG2-GRA1-ROP1) obtained by genetic engineering were...
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The Development of Toxoplasma gondii Recombinant Trivalent Chimeric Proteins as an Alternative to Toxoplasma Lysate Antigen (TLA) in Enzyme-Linked Immunosorbent Assay (ELISA) for the Detection of Immunoglobulin G (IgG) in Small Ruminants
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The Development of Toxoplasma gondii Recombinant Trivalent Chimeric Proteins as an Alternative to Toxoplasma Lysate Antigen (TLA) in Enzyme-Linked Immunosorbent Assay (ELISA) for the Detection of Immunoglobulin G (IgG) in Small Ruminants
PublikacjaThis study presents an evaluation of seventeen newly produced recombinant trivalent chimeric proteins (containing the same immunodominant fragment of SAG1 and SAG2 of Toxoplasma gondii antigens, and an additional immunodominant fragment of one of the parasite antigens, such as AMA1, GRA1, GRA2, GRA5, GRA6, GRA7, GRA9, LDH2, MAG1, MIC1, MIC3, P35, and ROP1) as a potential alternative to the whole-cell tachyzoite lysate (TLA) used...
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Characterization of recombinant homocitrate synthase from Candida albicans
PublikacjaLYS21 and LYS22 genes from Candida albicans encoding isoforms of homocitrate synthase (HCS), an enzyme catalyzing the first committed step in the L-lysine biosynthetic pathway, were cloned and expressed as NoligoHistagged fusion proteins in Escherichia coli. The purified gene products revealed HCS activity, i.e. catalyzed the condensation of α-ketoglutarate with acetyl-coenzyme A to yield homocitrate. The recombinant enzymes were purified...
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Protein & Cell
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PROTEIN JOURNAL
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PROTEUS
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The Role of Proteins in Food
PublikacjaThis chapter describes the effect of proteins on the sensory attributes and the biological value and safety of foods. The role of proteins depends on their amino acid composition and structure, on changes due to storage and processing, as well as on interactions with other food components. The effect on the sensory quality of foods is brought about by hydrophobicity, solubility, water holding capacity, gelling, film formation,...
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Use of MAG1 recombinant antigen for diagnosis of Toxoplasma gondii infection in humans
PublikacjaPraca opisuje klonowanie, oczyszczanie oraz zastosowanie w immunodiagnostyce toksoplazmozy antygenu rekombinantowego macierzy cyst tkankowych (MAG1) Toxoplasma gondii. Zastosowany system ekspresyjny pozwolił uzyskać dużą ilość rekombinantowego antygenu, który następnie wykorzystano w teście ELISA do wykrywania specyficzych przeciwciał anty-T. gondii klasy IgG w surowicach pacjentów chorych na toksoplazmozę. W przeprowadzonych badaniach...
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Toxoplasma gondii: Chimeric Dr fimbriae as a recombinant vaccine against toxoplasmosis
PublikacjaW publikacji przedstawiono protekcyjne i immunogenne właściowści chimerycznych fimbrii typu Dr zawierających sekwencje epitopowe antygenów SAG1, MAG1 oraz GRA1 Toxoplasma gondii. Uzyskane wyniki porównano z właściwościami immunogennymi antygenowych białek rekombinantowych oraz poliwalentnego antygenu natywnego. Antygeny rekombinantowe indukują silną odpowiedź immunologiczną i w 89% zapobiegają formowaniu się cyst w mózgu myszy....
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Toxoplasma gondii: an evaluation of diagnostic value of recombinant antigens in a murine model
PublikacjaPraca dotyczy oceny użyteczności sześciu rekombinantowych białek antygenowych Toxoplasma gondii (GRA1, GRA6, GRA7, p35, SAG1 i SAG2) w diagnozowaniu doświadczalnej toksoplazmozy u myszy. Przeprowadzone badania pozwoliły na określenie stopnia reaktywności zastosowanych antygenów z surowicami z wczesnej i przewlekłej toksoplazmozy. W przypadku ostrej fazy choroby obserwowano silną odpowiedź humoralną na antygeny GRA7, GRA6 i p35,...
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Toxoplasma gondii Recombinant Antigens in the Serodiagnosis of Toxoplasmosis in Domestic and Farm Animals
PublikacjaToxoplasmosis is caused by an intracellular protozoan, Toxoplasma gondii, and is a parasitic disease that occurs in all warm-blooded animals, including humans. Toxoplasmosis is one of the most common parasitic diseases of animals and results in reproductive losses. Toxoplasmosis in humans is usually caused by eating raw or undercooked meat or consuming dairy products containing the parasite. Diagnosis of toxoplasmosis is currently...
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PROTEIN SCIENCE
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Activity-based protein profiling guided identification of urine proteinase 3 activity in subclinical rejection after renal transplantation
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Cloning, expression, purification and characterization of recombinant trehalose synthase from Deinococcus radiodurans.
PublikacjaTrehalose (α-D-glucopyranosyl-1,1-α-D-glucopyranoside) is a nonreducing disacharide in which the two glucose molecules are linked trough a α-1,1-glycosidic bond. Trehalose is readily hydrolyzed to glucose and can be used as a reserve of that sugar in the cell. The presence of trehalose was found in the cells of fungi and yeasts, bacteria, nematodes, insects, eggs, pupae and some plants. The characteristics of trehalose make it...
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Production of Toxoplasma gondii MAG1 recombinant antigen in Leishmania tarentolae expression system
PublikacjaThe results of the work associated with the production of the Toxoplasma gondii matrix antigen 1 (MAG1) in the eukaryotic expression system of Leishmania tarentolae are shown.
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Properties of recombinant trehalose synthase from Deinococcus radiodurans expressed in Escherichia coli
PublikacjaA trehalose synthase gene from Deinococcus radiodurans(DSMZ 20539)containing 1659 bp reading frame encoding 552 amino acids was amplified using PCR. The gene was finally ligated into pET30Ek/LIC vector and expressed after isopropyl β-d-thiogalactopyranoside induction in Escherichia coli (DE3) Rosetta pLysS. The recombinanttrehalose synthase (DraTreS) containing a His6-tag at the C-terminus was purified by metal affinity chromatography...
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Efficient production of Staphylococcus simulans lysostaphin in a benchtop bioreactor by recombinant Escherichia coli.
PublikacjaLysostaphin is an enzyme with bactericidal activity against Staphylococcus aureus and other staphylococcal species. In spite of many advantages and promising results of preliminary research, the enzyme is still not widely used in medicine, veterinary or as a food preservative. One of the most important factors limiting application of the enzyme in clinical or technological practice is the high costs of its production. In the present...
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Testing the Diagnostic Utility of Recombinant Toxoplasma Gondii Chimeric Antigens – Generated Datasets
PublikacjaThe datasets titled Toxoplasma gondii recombinant chimeric antigens – IgM and IgG ELISAs – mouse serum samples and Toxoplasma gondii recombinant chimeric antigens – IgG and IgM ELISAs – human serum samples contain absorbance measurements obtained during serological tests using mouse and human sera in enzyme-linked immunosorbent assay (ELISA) tests based on recombinant chimeric antigens. The datasets allows a comparison of absorbance...
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The development of an indirect ELISA for the detection of goose parvovirus antibodies using specific VP3 subunits as the coating antigen
PublikacjaIn Poland, the leader in goose production in Europe, goose parovirus infection, or Derzsy’s disease (DD), must be reported to the veterinary administration due to the serious economic and epizootic threat to waterfowl production. Prophylactic treatment for DD includes attenuated live or inactivated vaccines. Moreover, the control of DD includes the monitoring of maternal derived antibody (MDA) levels in the offspring and antibody...
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Concentration of proteins in the whey stream
PublikacjaZastosowano metodę separacji pianowej do odzyskiwania białka z serwatki. Wykazano silny wpływ prędkości powietrza w kolumnie, początkowego stężenia białka w serwatce i pH roztworu na przebieg i skuteczność flotacji. Współczynnik wzbogacenia maleje ze wzrostem prędkości przepływu powietrza oraz ze wzrostem początkowego stężenia białka serwatkowego. Najwyższą wartość współczynnika wzbogacenia uzyskano dla pH naturalnej serwatki.
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Conserved motifs of MutL proteins
PublikacjatThe MutL protein is best known for its function in DNA mismatch repair (MMR). However, there isevidence to suggest that MutL is not only the linker connecting the functions of MutS and MutH in MMR,but that it also participates in other repair systems, such as Very Short Patch (VSP), Base Excision (BER)and Nucleotide Excision Repair (NER). This study set out to identify the most highly conserved aminoacid sequence motifs in MutL...
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Conservation and diversity of MutS proteins
PublikacjaThe homologues of MutS, mismatch repair protein, exist in all prokaryotes, with the exception of Actinobacteria, Mollicutes and part of the Archaea. Multiple alignments of 316 MutS amino acid sequences from 169 species revealed conserved residues and sequence motifs distinguishing MutS homologues. All MutS homologues show high conservation within the ATPase domain. MutS1, the homologue responsible for DNA mismatch recognition,...
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ADVANCES IN PROTEIN CHEMISTRY
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JOURNAL OF PROTEIN CHEMISTRY
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A method for the production of D-tagatose using a recombinant Pichia pastoris strain secreting beta-D-galactosidase from Arthrobacter chlorophenolicus and a recombinant L-arabinose isomerase from Arthrobacter sp. 22c
PublikacjaD-Tagatose is a natural monosaccharide which can be used as a low-calorie sugar substitute in food, beverages and pharmaceutical products. It is also currently being tested as an anti-diabetic and obesity control drug. D-Tagatose is a rare sugar, but it can be manufactured by the chemical or enzymatic isomerization of D-galactose obtained by a beta-D-galactosidase-catalyzed hydrolysis of milk sugar lactose and the separation of...
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Protein overproduction alters exosome secretion in Chinese hamster ovary cells
PublikacjaDespite the abundance of available cell lines, nearly 70% of all recombinant therapeutic proteins today are produced in Chi-nese hamster ovary (CHO) cells. The impact of protein overproduction on the secretion of exosomes by CHO cells has been investigated here. Increased secretion of extracellular vesicles (EVs) by protein overexpressing CHO cells was demonstrated with protein content assay, nanoparticle tracking analysis, and...
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Potential diagnostic utility of recombinant Toxoplasma gondii proteins for detecting anti-Toxoplasma gondii IgG antibodies in horses serum samples
Dane BadawczeToxoplasmosis is caused by an intracellular protozoan, Toxoplasma gondii, and is a parasitic disease that occurs in all warm-blooded animals, including humans. Toxoplasmosis is one of the most common parasitic diseases of animals and results in reproductive losses. Diagnosis of toxoplasmosis is currently based on serological assays using native antigens...
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Potential diagnostic utility of recombinant Toxoplasma gondii proteins for detecting anti-Toxoplasma gondii IgG antibodies in goat serum samples
Dane BadawczeToxoplasmosis is caused by an intracellular protozoan, Toxoplasma gondii, and is a parasitic disease that occurs in all warm-blooded animals, including humans. Toxoplasmosis is one of the most common parasitic diseases of animals and results in reproductive losses. Diagnosis of toxoplasmosis is currently based on serological assays using native antigens...
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Potential diagnostic utility of recombinant Toxoplasma gondii proteins for detecting anti-Toxoplasma gondii IgG antibodies in feline serum samples
Dane BadawczeToxoplasmosis is caused by an intracellular protozoan, Toxoplasma gondii, and is a parasitic disease that occurs in all warm-blooded animals, including humans. Toxoplasmosis is one of the most common parasitic diseases of animals and results in reproductive losses. Diagnosis of toxoplasmosis is currently based on serological assays using native antigens...
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Toxoplasma gondii recombinant antigens as tools for serodiagnosis of human toxoplasmosis: current status of studies
PublikacjaToxoplasma gondii is a parasitic protozoan which is the cause of toxoplasmosis. Although human toxoplasmosis in healthy adults is usually asymptomatic, serious disease can occur in the case of congenital infections and immunocompromised individuals. Furthermore, despite the exact recognition of its etiology, it still presents a diagnostic problem. Diagnosis of toxoplasmosis is mainly based on the results of serological tests detecting...
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Use of Escherichia coli Nissle 1917 producing recombinant colicins for treatment of IBD patients
PublikacjaPatients with Crohn’s Disease and ulcerative colitis infected with Adherent-Invasive Escherichia coli strains constitute the largest group among Inflammatory Bowel Disease subjects, when taking into account all known etiological agents of the disease. A possible link between these pathogenic bacteria and inflammation process has gained the confidence in recently published papers. Observed enteric neuroglial cells apoptosis and...