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Search results for: ESCHERICHIA COLI EXPRESSION SYSTEM
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Novel thermostable single-stranded DNA-binding protein (SSB) from Deinococcus geothermalis
PublicationW celu zbadania właściwości biochemicznych białka wiążącego się do jednoniciowego DNA (SSB) pochodzącego z Deinococcus geothermalis (DgeSSB), sklonowano gen ssb otrzymany w reakcji PCR, a następnie uzyskano wydajną biosyntezę białka DgeSSB. Gen składa się z 900 par zasad, kodujących białko złożone z 300 reszt aminokwasowych, o wyliczonej masie molekularnej równej 32.45 kDa. Sekwencja aminokwasowa wykazuje 43, 44% i 75% identyczności...
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Identification and properties of the Deinococcus grandis andDeinococcus proteolyticus single-stranded DNA binding proteins (SSB)
PublicationW celu poznania właściwości biochemicznych białek SSB pochodzących z Deinococcus grandis (DgrSSB) i Deinococcus proteolyticus (DprSSB) sklonowano zaamplifikowane w reakcji PCR geny ssb w systemie ekspresyjnym Escherichia coli. Geny składają się z 891(DgrSSB) i 876 (DprSSB) nukleotydów kodujących odpowiednio 296 i 291 reszt aminokwasowych o teoretycznie wyznaczonej masie cząsteczkowej dla białek, równej 32,29 i 31,33 kDa. Sekwencja...
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Novel single-stranded DNA-binding proteins from extreme psychrophilic bacterium Psychromonas ingrahamii 37
PublicationWe report the identification and characterization of the single-stranded DNA binding protein (SSB) from extreme psychrophilic bacterium Psychromonas ingrahamii 37 (PinSSB) that grows expotentially at -12°C and may well grow at even lower temperatures. PinSSB is one of the largest known bacterial SSB protein consisting 222 amino acid residues with a calculated molecular mass of 25.1 kDa. The analysis by gel filtration chromatography...
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Characterization of exceptionally thermostable single-stranded DNA-binding proteins from Thermotoga maritima and Thermotoga neapolitana
PublicationBACKGROUND: In recent years, there has been an increasing interest in SSBs because they find numerous applications in diverse molecular biology and analytical methods.RESULTS: We report the characterization of single-stranded DNA binding proteins (SSBs) from the thermophilic bacteria Thermotoga maritima (TmaSSB) and Thermotoga neapolitana (TneSSB). They are the smallest known bacterial SSB proteins, consisting of 141 and 142 amino...
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Źródła, otrzymywanie i przydatność niektórych termostabilnych enzymów w przetwórstwie skrobi i mleka
PublicationPrezentowane wyniki wskazują na możliwość usprawnienia produkcji syropów skrobiowych przez zastosowanie rekombinantowej alfa-amylazy z Pyrococcus woesei i termostabilnej alfa-glukozydazy z Thermus thermophilus. Wykazano także przydatność rekombinantowej, termosta-bilnej beta-galaktozydazy z Pyrococcus woesei do wytwarzania bez-laktozowego mleka i serwatki. Enzym ten wykazuje dużą termostabil-ność i znaczną aktywność w szerokim...
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Correlation between the number of Pro-Ala repeats in the EmrA homologue of Acinetobacter baumannii and resistance to netilmicin, tobramycin, imipenem and ceftazidime
PublicationAcinetobacter baumannii coccobacilli are dangerous to patients in intensive care units because of their multidrug resistance to antibiotics, developed mainly in the past decade. This study aimed to examine whether there is a significant correlation between the number of Pro-Ala repeats in the CAP01997 protein, the EmrA homologue of A. baumannii, and resistance to antibiotics. A total of 79 multidrug-resistant A. baumannii strains...
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The New LM-PCR/Shifter Method for the Genotyping of Microorganism
PublicationTechniques relies on the ligation of appropriates adapters (LM-PCR) as AFLP, PCR MP and ADSRRS are successfully used for epidemiological studies for prokaryotic and eukaryotic microorganisms. In this study we propose a new method, called the LM-PCR/Shifter, based on the use of a Class IIS restriction enzyme giving restriction fragments with different 4 base 5' overhangs (Shifter) and the ligation of appropriate oligonucleotide...
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Antifungal Effect of Penicillamine Due to the Selective Targeting of L-Homoserine O-Acetyltransferase
PublicationDue to the apparent similarity of fungal and mammalian metabolic pathways, the number of established antifungal targets is low, and the identification of novel ones is highly desirable. The results of our studies, presented in this work, indicate that the fungal biosynthetic pathway of L-methionine, an amino acid essential for humans, seems to be an attractive perspective. The MET2 gene from Candida albicans encoding L-homoserine...
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Encapsulation of Amikacin into Microparticles Based on Low-Molecular-Weight Poly(lactic acid) and Poly(lactic acid-co-polyethylene glycol)
PublicationThe aim of this study was to fabricate novel microparticles (MPs) for efficient and long-term delivery of amikacin (AMI). The emulsification method proposed for encapsulating AMI employed low-molecular-weight poly(lactic acid) (PLA) and poly(lactic acid-co-polyethylene glycol) (PLA−PEG), both supplemented with poly(vinyl alcohol) (PVA). The diameters of the particles obtained were determined as less than 30 μm. Based on an in-vitro...
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Polyurethane porous scaffolds (PPS) for soft tissue regenerative medicine applications
PublicationTissue engineering requires suitable polymeric scaffolds, which act as a physical support for regenerated tissue. A promising candidate might be polyurethane (PUR) scaffold, due to the ease of the PUR properties design, which can be adjusted directly to the intended purpose. In this study, we report a successful fabrication of porous polyurethane scaffolds (PPS) using solvent casting/particulate leaching technique combined with...
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Synteza nowych aminokwasowych i peptydowych analogów kwasu mykofenolowego oraz ich aktywność biologiczna i mikrobiologiczna
PublicationKwas mykofenolowy (MPA) oraz jego dwie stosowane klinicznie pochodne: mykofenolan sodu (MPS, CellCept) i mykofenolan mofetylu (MMF, Myfortic) są jednymi z najczęściej używanych leków hamujących proliferację limfocytów. Obok aktywności immunosupresyjnej, farmaceutyk wykazuje również właściwości przeciwbakteryjne, przeciwgrzybicze, przeciwwirusowe a także przeciwnowotworowe. W ramach realizacji swojej pracy doktorskiej zaprojektowałam...
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Bee Pollen and Bee Bread as a Source of Bacteria Producing Antimicrobials
PublicationThe principal objective of the study was the isolation and identification of bacteria that are present in mature bee bread (BB) and dried (ready for selling and consumption) bee pollen (BP). Obtained isolates were screened for their potential to inhibit select human pathogenic bacteria and their ability to produce enzymes of particular industrial importance. Four and five samples of BP and BB, respectively, were used for the study....
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Production of Toxoplasma gondii MAG1 recombinant antigen in Leishmania tarentolae expression system
PublicationThe 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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Defected Ag/Cu-MOF as a modifier of polyethersulfone membranes for enhancing permeability, antifouling properties and heavy metal and dye pollutant removal
PublicationIn this study, a novel bimetallic metal-organic framework (MOF) i.e. Ag/Cu-MOF was synthesized using a solvothermal method and later incorporated at different concentrations (0.1–2 wt%) using a phase inversion method for modification and antifouling property improvement of polyethersulfone (PES) membranes. The resulting Ag/Cu-MOF characteristics were investigated using different techniques, such as FTIR, XRD, FE-SEM and EDX. The...
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A new MIC1-MAG1 recombinant chimeric antigen can be used instead of the Toxoplasma lysate antigen in the serodiagnosis of human toxoplasmosis
PublicationW niniejszej publikacji po raz pierwszy przedstawiono otrzymanie i oszacowanie przydatności diagnostycznej nowego chimerycznego antygenu MIC1-MAG1 Toxoplasma gondii do rozpoznawania toksoplazmozy u ludzi. Antygen chimeryczny MIC1-MAG1 został otrzymany jako białko fuzyjne zawierające dwie domeny oligohistydynowe na N- i C-końcu z użyciem systemu ekspresyjnego Escherichia coli. Po oczyszczeniu z wykorzystaniem metody chromatografii...
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Characterization of recombinant homocitrate synthase from Candida albicans
PublicationLYS21 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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Electrochemical detection of bacterial endotoxin lipopolysaccharide (LPS) on gold electrode modified with DAL-PEG-DK5-PEG-OH - Antimicrobial peptide conjugate
PublicationThis work describes fabrication of gold electrodes modified with peptide conjugate DAL-PEG-DK5-PEG-OH that enables ultra-sensitive detection of lipopolysaccharide (LPS) isolated from the reference strain of Escherichia coli O26:B6. The initial step of the established procedure implies immobilization of the fully protected DAL-PEG-DK5-PEG-OH peptide on the surface of the gold electrode previously modified by cysteamine. Then side...
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Systemy ekspresji genów - Wykład - 2021_22
e-Learning CoursesTematyka wykładów kursu przedstawia następujące treści: Ekspresja genów w organizmach żywych. Źródła informacji:gdy sekwencja genu jest znana, gdy sekwencja genu nie jest znana, bazy danych, biblioteki DNA genomowego, biblioteki cDNA, narzędzia on-line używane do analizy sekwencji DNA. System ekspresyjny genów - pojęcia podstawowe. Kryteria wyboru systemu ekspresyjnego do produkcji białek heterogenicznych w zależności od ich...
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Zaawansowane metody charakterystyki i uzdatniania wody w basenach kąpielowych
PublicationW literaturze przedmiotu szeroko opisywane są zagrożenia dla zdrowia kąpiących się i obsługi ze względu na losowe występowanie w wodzie bakterii grupy coli, bakterii Escherichia coli typu kałowego, gronkowca koagulazo-dodatniego, pierwotniaków Cryptosporidium Parvum, Gardia i innych mikroorganizmów, a także produktów reakcji związków dezynfekujących z zanieczyszczeniami organicznymi w wodzie. Stosowane dotychczas technologie uzdatniania...
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Identification and cloning of C. albicans SC5314 genes encoding L-methionine biosynthetic pathway enzymes.
Open Research DataEnzymes of fungal L-methionine biosynthetic pathway: homoserine O-acetyltransferase (Met2p), O-acetylhomoserine sulfhydrylase (Met15p) and cystathionine-γ-synthase (Str2p) could be exploited as molecular targets for antifungal chemotherapy. The goal of the study was to identify and clone genes encoding mentioned above enzymes. MET2, MET15 and STR2 genes...
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Beata Krawczyk dr hab.
PeopleBeata Krawczyk, PhD, Professor Academic degrees, education, qualifications: B. Krawczyk earned his master degree in Biology at the University of Gdansk, Biology Faculty, in 1986, and her PhD in Molecular Biology at the University of Gdansk, Biology Faculty, in 1996. Postdoctoral degree (habilitation) in biological sciences in the discipline biology, Faculty of Natural Sciences, University of Szczecin, Szczecin, in 2009. In 2012...
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Sposób otrzymywania rekombinantowego szczepu Escherichia coli Rosetta (DE3) pLysS oraz sposób wytwarzania białka syntazy trehalozy Deinococcus radiodurans DSMZ 20539
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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
PublicationTrzy 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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Structural studies of a cold-adapted dimeric Beta-D-galactosidase from Paracoccus sp. 32d
PublicationThe crystal structure of a novel dimeric [beta]-D-galactosidase from Paracoccus sp. 32d (Par[beta]DG) was solved in space group P212121 at a resolution of 2.4 Å by molecular replacement with multiple models using the BALBES software. This enzyme belongs to glycoside hydrolase family 2 (GH2), similar to the tetrameric and hexameric [beta]-D-galactosidases from Escherichia coli and Arthrobacter sp. C2-2, respectively. It is the second...
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Bioactivation of Konjac Glucomannan Films by Tannic Acid and Gluconolactone Addition
PublicationWound healing is a dynamic process that requires an optimal extracellular environment, as well as an accurate synchronization between various cell types. Over the past few years, great efforts have been devoted to developing novel approaches for treating and managing burn injuries, sepsis, and chronic or accidental skin injuries. Multifunctional smart-polymer-based dressings represent a promising approach to support natural healing...
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Lipopolysaccharides: regulated biosynthesis and structural diversity
PublicationThe cell envelope of Gram-negative bacteria contains two distinct membranes, an inner (IM) and an outer (OM) membrane, separated by the periplasm, a hydrophilic compartment that includes a thin layer of peptidoglycan. The most distinguishing feature of such bacteria is the presence of an asymmetric OM with phospholipids located in the inner leaflet and lipopolysaccharides (LPSs) facing the outer leaflet. The maintenance of this...
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Józef Kur prof. dr hab.
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The database of localization and expression of classical aquaporins (AQPs) in the male reproductive system in cattle
Open Research DataThe data present research results being part of the OPUS-22 project entitled “In search of new markers of male fertility in cattle. Aquaporins expression in the reproductive organs and sperm of the bulls (Bos taurus)” financed by the National Science Center in Poland (grant no. 2021/43/B/NZ9/00204). The aim of this part of the project was (i) to determine...
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Preclusion of irreversible destruction of Dr adhesin structures by a high activation barrier for the unfolding stage of the fimbrial DraE subunit
PublicationFimbrie Dr uropatogennych szczepów Escherichia coli stanowią przykład eksponowanych powierzchniowo struktur adhezyjnych podlegających biogenezie według zakonserwowanego, pośród bakterii Gram-ujemnych, systemu "chaperone-usher" ("białko opiekuńcze-zewnątrzbłonowe białko kanałowe"). Powyższe struktury adhezyjne są niezbędne w procesie specyficznej adhezji bakterii do receptorów zlokalizowanych na powierzchni docelowych komórek gospodarza....
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Possibility of Using Tea Fungus for Fermentation of Beetroot and Carrot Pomace Beverages
PublicationFermented beverages obtained from plant-based raw materials are becoming more popular due to their beneficial effects on human health. These include kombucha beverages, which are obtained by fermenting tea brew using the so-called tea fungus, which includes acetic acid bacteria and osmophilic yeast. A consortium of these microorganisms could also be used to prepare functional fermented beverages obtained from extracts of vegetable...
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FUNKCJONALIZOWANIE 2H-TIAZOLO[3,2-α]PIRYDYN-5-(3H)-ONÓW, ALTERNATYWNA ŚCIEŻKA SYNTEZY NOWYCH PILICYDÓW
PublicationUropatogenne szczepy Escherichia coli są przyczyną blisko 80% przypadków zakażeń dróg moczowych (ang. UTI- Urinary tract infection). Ponadto, aż 30% odmiedniczkowego zapalenia nerek występującego u ciężarnych kobiet jest spowodowane przez szczepy E.coli Dr+. Szerząca się lekooporność bakterii i trudności w leczeniu UTI wymusiły na środowiskach naukowych poszukiwania nowych związków chemicznych pełniących rolę chemoterapeutyków. Na...
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Fusion of Taq DNA polymerase with single-stranded DNA binding-like protein of Nanoarchaeum equitans—Expression and characterization
PublicationDNA polymerases are present in all organisms and are important enzymes that synthesise DNA molecules. They are used in various fields of science, predominantly as essential components for in vitro DNA syntheses, known as PCR. Modern diagnostics, molecular biology and genetic engineering need DNA polymerases which demonstrate improved performance. This study was aimed at obtaining a new NeqSSB-TaqS fusion DNA polymerase from the...
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A New Expression System Based on Psychrotolerant Debaryomyces macquariensis Yeast and Its Application to the Production of Cold-Active β-D-Galactosidase from Paracoccus sp. 32d
PublicationYeasts provide attractive host/vector systems for heterologous gene expression. The currently used yeast-based expression platforms include mesophilic and thermotolerant species. A eukaryotic expression system working at low temperatures could be particularly useful for the production of thermolabile proteins and proteins that tend to form insoluble aggregates. For this purpose, an expression system based on an Antarctic psychrotolerant...
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Evaluation of antimicrobial activity of porous composites based on chitosan/poly (vinyl alcohol)
Open Research DataThe dataset contains the results of microbiological tests of composite porous materials whose activity was assessed for their ability to reduce the number of Escherichia coli and Staphylococcus aureus strain, representing the Gram (-) and Gram (+) bacteria, respectively.
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Structural analogues of reactive intermediates as inhibitors of glucosamine-6-phosphate synthase and phosphoglucose isomerase
PublicationCentra aktywne izomerazy fosfoglukozowej (PGI) oraz domeny izomerazowej (HPI) syntazy glukozamino-6-fosforanu (GlcN-6-P), wykazują podobieństwo ułożenia przestrzennego kluczowych reszt aminokwasowych, z wyjątkiem reszty Arg272 PGI i reszt Lys603 i Lys485 HPI. Dziesięć pochodnych D-heksitolo-6-P, kwasu 5-fosfoarabonowego i kwasu 6-fosfoglukonowego, strukturalnych analogów cis-enolaminy lub cis-enolanu, przypuszczalnych stanów przejściowych...
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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
PublicationD-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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The database of localization and expression of aquaporin 3 (AQP3), aquaporin 7 (AQP7) and aquaporin 9 (AQP9) in the male reproductive system in cattle. Morphometric studies. Localization of zinc finger transcription factor GATA-4.
Open Research DataThe data present research results being part of the OPUS-22 project entitled “In search of new markers of male fertility in cattle. Aquaporins expression in the reproductive organs and sperm of the bulls (Bos taurus)” obtained from the National Science Center in Poland (grant no. 2021/43/B/NZ9/00204). The aim of this part of the project was (i) to determine...
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Expression profile of messenger and micro RNAs related to the histaminergic system in patients with five subtypes of breast cancer
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Detection of cellulose production capacity of recombinant Escherichia coli strains BL21(DE3) and AAEC191A
Open Research DataBacteria that form biofilms generate an extracellular matrix (ECM), where cellulose stands out as a key constituent. An approach for assessing microorganisms' cellulose production involves using calcofluor white staining on colonies. In this method, a fluorescent dye (calcofluor-white) is introduced to a stable YESCA substrate composed of casamino acids,...
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Detection of cellulose production capacity of recombinant Escherichia coli strains BL21(DE3) and AAEC191A
Open Research DataBacteria that form biofilms generate an extracellular matrix (ECM), where cellulose stands out as a key constituent. An approach for assessing microorganisms' cellulose production involves using calcofluor white staining on colonies. In this method, a fluorescent dye (calcofluor-white) is introduced to a stable YESCA substrate composed of casamino acids,...
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Detection of cellulose production capacity of recombinant Escherichia coli strains BL21(DE3) and AAEC191A
Open Research DataBacteria that form biofilms generate an extracellular matrix (ECM), where cellulose stands out as a key constituent. An approach for assessing microorganisms' cellulose production involves using calcofluor white staining on colonies. In this method, a fluorescent dye (calcofluor-white) is introduced to a stable YESCA substrate composed of casamino acids,...
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Detection of cellulose production capacity of recombinant Escherichia coli strains BL21(DE3) and AAEC191A
Open Research DataBacteria that form biofilms generate an extracellular matrix (ECM), where cellulose stands out as a key constituent. An approach for assessing microorganisms' cellulose production involves using calcofluor white staining on colonies. In this method, a fluorescent dye (calcofluor-white) is introduced to a stable YESCA substrate composed of casamino acids,...
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Gracjana Klein-Raina dr hab.
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Sekwencja DNA, startery oligonukleotydowe, sekwencja DNA wraz z miejscami rozpoznania dla enzymów restrykcyjnych NcoI i HindIII, wektor ekspresyjny, rekombinowany szczep Escherichia coli TOP10/F' i jego sposób otrzymywania oraz sposób wytwarzania termostabilnego białka SSB-podobnego Deinococcus radiopugnans DSMZ 12027
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Sekwencja DNA, startery oligonukleotydowe, sekwencja DNA wraz z miejscami rozpoznania dla enzymów restrykcyjnych NcoI i HindIII, wektor ekspresyjny, rekombinowany szczep Escherichia coli TOP10/F' i jego sposób otrzymywania oraz sposób wytwarzania termostabilnego białka SSB-podobnego Deinococcus radiopugnans DSMZ 11303
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Study of the influence of the presence of Dr fimbriae on the hydrophobicity of cells of recombinant Escherichia coli strains: AAEC191A and BL21(DE3)
Open Research DataThe ability of a cell surface to repel or adhere to surfaces is crucial, and it was assessed through a microbiological adhesion test involving hexadecane and xylene. In this method, microorganisms in an aqueous layer are mixed with an organic layer, and after completion, the absorbance of each sample is measured at a 570 nm wavelength. The results obtained...
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The effect of oxidized cholesterol on barrier functions and IL-10 mRNA expression in human intestinal epithelium co-cultured with dendritic cells in the transwell system
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Aravind Ayyolath
PeopleI am interested in the biology of bacterial stress response and its growth under stress conditions. Our lab focuses on two aspects; protein folding and lipopolysaccharide biosynthesis and we address them with Escherichia coli models. Our recent studies have provided more evidence regarding several genes involved in the survival of bacteria under stress. I am also interested in disease biology and also in identifying proper therapeutic...
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Rafał Piątek dr hab. inż.
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Beata Zalewska-Piątek dr hab.
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