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Search results for: ESCHERICHIA COLI EXPRESSION SYSTEM
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A two-plasmid Escherichia coli system for expression of Dr adhesins
PublicationPraca opisuje konstrukcję wydajnego systemu produkcji fimbrii Dr. System składa się z dwóch plazmidów. Plazmid pACYCpBAD-DraC-C-His zawiera pod kontrolą promotora arabinozowego gen draC kodujący białko kanałotwórcze DraC. Plazmid pET-30b-sygDraBE zawiera pod kontrolą promotora T7lac geny draB i draE kodujące odpowiednio białko opiekuńcze DraB oraz adhezynę DraE. Oba plzamidy posiadają różne ori replikacji co pozwala na ich jednoczesne...
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Influence of Escherichia coli on Expression of Selected Human Drug Addiction Genes
PublicationThe impact of enteric microflora on the expression of genes associated with cocaine and amphetamine addiction was described. Human genome-wide experiments on RNA transcripts expressed in response to three selected Escherichia coli strains allowed for significant alteration (p > 0.05) of the linear regression model between HT-29 RNA transcripts associated with the KEGG pathway:hsa05030:Cocaine addiction after 3 h stimulation with...
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Expression of goose parvovirus whole VP3 protein and its epitopes in Escherichia coli cells
PublicationThe aim of this study was the expression of goose parvovirus capsid protein (VP3) and its epitopes in Escherichia coli cells. Expression of the whole VP3 protein provided an insufficient amount of protein. In contrast, the expression of two VP3 epitopes (VP3ep4, VP3ep6) in E. coli, resulted in very high expression levels. This may suggest that smaller parts of the GPV antigenic determinants are more efficiently expressed than the...
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Expression of Deinococcus geothermalis Trehalose Synthase Gene in Escherichia coli and Its Enzymatic Properties
PublicationA novel trehalose synthase gene from Deinococcus geothermalis (DSMZ 11300) containing 1,692 bp reading-frame encoding 564 amino acids was amplified using PCR. The gene was ligated into pET30Ek/LIC vector and expressed after isopropyl alfa-D-thiogalactopyranoside induction in Escherichia coli BL21(DE3)pLysS. The recombinant trehalose synthase (DgeoTreS) containing a His6 tag at the C-terminus was purified by metal affinity chromatography...
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Urinary Tract Infections of Escherichia coli Strains of Chaperone-Usher System
PublicationZakażenia dróg moczowych stanowią bardzo poważny problem zdrowotny i ekonomiczny dotyczący milionów ludzi, każdego roku na całym świecie. Najbardziej rozpowszechnionym czynnikiem etiologicznym tego typu infekcji bakteryjnych, obejmujących górne i dolne drogi moczowe, są szczepy E. coli, reprezentujące około 80% przypadków. Uropatogenne szczepy E. coli produkują kilka czynników urowirulentncyh, które można podzielić na dwa główne...
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A freeze-thaw method for desintegration of Escherichia coli cells producing T7 lysozyme used in pBAD expression systems
PublicationPlazmid pLysN zawierający gen kodujący lizozym T7 pod kontrolą promotora lac został skonstruowany w celu ułatwienia dezintegracji komórek po ekspresji rekombinantowych białek w systemach ekspresji indukowanych arabinozą. Użyteczność plazmidu została przetestowana w komórkach Escherichia coli TOP10 i E. coli LMG194, niosących plazmid pBADMHADgeSSB, zawierający gen kodujący białko SSB Deinococcus geothermalis pod kontrolą promotora...
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High yield expression and single step purification of Toxoplasma gondii SAG1, GRA1, GRA7 antigens in Escherichia coli.
PublicationPraca przedstawia metodę uzyskiwania antygenów rekombinowanych Toxoplasmagondii o wysokiej czystości, które można zastosować w immunodiagnostyce. Antygeny T. gondii eksprymowane w komórkach E. coli zawierały polihistydynowedomeny fuzyjne na końcu N i C białka. Pozwalało to na jednoetapowe oczyszczanie białek metodą chromatografii metalopowinowactwa na złożu Ni2+ -IDA-Sepharose. Immunoreaktywność antygenów rekombinowanych...
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Escherichia coli lipopolysaccharide may affect the endothelial barrier and IL‐10 expression of apolipoprotein B100‐pulsed dendritic cells
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Type II secretory pathway for surface secretion of DraD invasin from the uropathogenic Escherichia coli Dr+ strain
PublicationWłaściwości wirulentne uropatogennych szczepów Escherichia coli Dr+ IH11128 związane są z obecnością struktur fimbrialnych typu Dr i białka inwazyny DraD, które może na powierzchni komórki bakteryjnej pełnić rolę fimbrialnej podjednostki szczytowej. Ostatnie badania wykazały, że białko DraD jest eksponowane na powierzchni bakterii w dwóch formach: związanej ze strukturą fimbrialną i jako niezależna od fimbrii otoczka białkowa....
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Extracellular secretion of Pseudoalteromonas sp. cold-adapted esterase in Escherichia coli in the presence of Pseudoalteromonas sp. components of ABC transport system
PublicationEnzymy lipolityczne, głównie lipazy i esterazy, stanowią cenną grupę biokatalizatorów wykorzystywanych w przemyśle. Coraz większe zainteresowanie tą klasą enzymów sprawiło, że poszukuje się alternatywnych źródeł ich pozyskiwania. W celu zbadania wpływu białek transportu ABC Pseudoalteromonas sp. 643A na ekspresję genu, sekrecję i aktywność esterazy EstA z tego mikroorganizmu w systemie ekspresyjnym E. coli skonstruowano układ do...
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Study of the influence of the presence of Dr fimbriae on the adherence of cells of the recombinant Escherichia coli strain: AAEC191A/pACYCpBAD to polystyrene in a dynamic system
Open Research DataThe attachment of bacteria begins the process of surface colonization, called biofilm development, characterized by a number of physicochemical and molecular interactions. Adherence to inert surfaces typically involves nonspecific interactions, whereas adherence to biological surfaces is associated with specific ligand-receptor interactions. In this...
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Study of the influence of the presence of Dr fimbriae on the adherence of cells of the recombinant Escherichia coli strain: AAEC191A/pACYCpBAD to glass in a dynamic system
Open Research DataThe attachment of bacteria begins the process of surface colonization, called biofilm development, characterized by a number of physicochemical and molecular interactions. Adherence to inert surfaces typically involves nonspecific interactions, whereas adherence to biological surfaces is associated with specific ligand-receptor interactions. In this...
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Study of the influence of the presence of Dr fimbriae on the adherence of cells of the recombinant Escherichia coli strain: AAEC191A/pCC90 to glass in a dynamic system
Open Research DataThe attachment of bacteria begins the process of surface colonization, called biofilm development, characterized by a number of physicochemical and molecular interactions. Adherence to inert surfaces typically involves nonspecific interactions, whereas adherence to biological surfaces is associated with specific ligand-receptor interactions. In this...
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Study of the influence of the presence of Dr fimbriae on the adherence of cells of the recombinant Escherichia coli strain: AAEC191A/pCC90 to polystyrene in a dynamic system
Open Research DataThe attachment of bacteria begins the process of surface colonization, called biofilm development, characterized by a number of physicochemical and molecular interactions. Adherence to inert surfaces typically involves nonspecific interactions, whereas adherence to biological surfaces is associated with specific ligand-receptor interactions. In this...
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Study of the influence of the presence of Dr fimbriae on the adherence of cells of the recombinant Escherichia coli strain: BL21(DE3)/pCC90 to polystyrene in a dynamic system
Open Research DataThe attachment of bacteria begins the process of surface colonization, called biofilm development, characterized by a number of physicochemical and molecular interactions. Adherence to inert surfaces typically involves nonspecific interactions, whereas adherence to biological surfaces is associated with specific ligand-receptor interactions. In this...
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Study of the influence of the presence of Dr fimbriae on the adherence of cells of the recombinant Escherichia coli strain: BL21(DE3)/pACYCpBAD to glass in a dynamic system
Open Research DataThe attachment of bacteria begins the process of surface colonization, called biofilm development, characterized by a number of physicochemical and molecular interactions. Adherence to inert surfaces typically involves nonspecific interactions, whereas adherence to biological surfaces is associated with specific ligand-receptor interactions. In this...
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Study of the influence of the presence of Dr fimbriae on the adherence of cells of the recombinant Escherichia coli strain: BL21(DE3)/pCC90 to glass in a dynamic system
Open Research DataThe attachment of bacteria begins the process of surface colonization, called biofilm development, characterized by a number of physicochemical and molecular interactions. Adherence to inert surfaces typically involves nonspecific interactions, whereas adherence to biological surfaces is associated with specific ligand-receptor interactions. In this...
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Study of the influence of the presence of Dr fimbriae on the adherence of cells of the recombinant Escherichia coli strain: BL21(DE3)/pACYCpBAD to polystyrene in a dynamic system
Open Research DataThe attachment of bacteria begins the process of surface colonization, called biofilm development, characterized by a number of physicochemical and molecular interactions. Adherence to inert surfaces typically involves nonspecific interactions, whereas adherence to biological surfaces is associated with specific ligand-receptor interactions. In this...
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Efficient production of Staphylococcus simulans lysostaphin in a benchtop bioreactor by recombinant Escherichia coli.
PublicationLysostaphin 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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Comparative analysis of mRNA transcripts of HT-29 cell line expressed in identical quantities for pathogenic E. coli strains UM146 and UM147 with control Escherichia coli Nissle 1917
PublicationAim of study was comparative analysis of mRNA transcripts of HT-29 cell line, expressed in identical quantities for the combination of pathogenic and non-pathogenic Escherichia coli strains. HT-29 confluent monolayers infection with two pathogenic E. coli strains UM146 and UM147 resulted in two sets of mRNA transcripts that were identical with RNA transcripts obtained for non-pathogenic one strain E. coli Nissle 1917. In this study...
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New Approaches for Escherichia coli Genotyping
PublicationEasy-to-perform, fast, and inexpensive methods of differentiation of Escherichia coli strains beyond the species level are highly required. Herein two new, original tools for genotyping of E. coli isolates are proposed. The first of the developed method, a PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) test uses a highly variable fliC gene, encoding the H antigen as a molecular target. The designing...
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Enzymatic degradation and transport of endothiopeptides into Escherichia coli K12 mutant strains
PublicationZbadano transport trzech otrzymanych na drodze syntezy ditiopeptydów do komórek Escherichia coli K12. Ponadto zbadano szybkość hydrolizy tych związków. Wszystkie peptydy są transportowane przez permeazy, natomiast są bardziej oporne na hydrolizę enzymatyczną w stosunku do naturalnych peptydów.
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Multiple transcriptional factors regulate transcription of the rpoE gene in Escherichia coli under different growth conditions and when the lipopolysaccharide biosynthesis is defective.
PublicationThe RpoE sigma factor is essential for the viability of Escherichia coli. RpoE regulates extracytoplasmic functions including lipopolysaccharide (LPS) translocation and some of its non-stoichiometric modifications. Transcription of the rpoE gene is positively autoregulated by EσE and by unknown mechanisms that control the expression of its distally located promoter(s). Mapping of 5′ ends of rpoE mRNA identified five new transcriptional...
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Escherichia coli - zapomniany patogen posocznic
PublicationPrzedstawiono próbę retrospektywnej analizy częstości występowania bakteriemii Gram-ujemnych, wywołanych przez E. coli u chorych hospitalizowanych w ośmiu grupach oddziałów klinicznych SPSK1 ACK AM w Gdańsku w latach 2002-2004. Dokonano oceny częstości występowania bakteriemii o etologii E. coli, liczby zgonów związanych bezpośrednio z posocznicą, analizy potencjalnych wrót zakażenia oraz analizy lekowrażliwości szczepów E. coli...
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Phosphorylation‐mediated regulation of heat shock response in Escherichia coli
PublicationEscherichia coli has two heat shock regulons under the transcriptional control of Esigma(32) and Esigma(E) RNA polymerases. These polymerases control the expression of genes, the products of which are needed for correct folding of proteins in the cytoplasm and the extracytoplasm respectively. In this study, we report that mutations in a tyrosine phosphatase-encoding gene led to decreased activity of these heat shock regulons. The...
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Biochemical characteristic of biofilm of uropathogenic Escherichia coli Dr+ strains.
PublicationUrinary tract infections caused by Escherichia coli are very common health problem in the developed countries. The virulence of the uropathogenic E. coli Dr+ IH11128 is determined by Dr fimbriae, which are homopolymeric structures composed of DraE subunits with the DraD protein capping the fiber. In this study, we have analyzed the structural and biochemical properties of biofilms developed by E. coli strains expressing Dr fimbriae...
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A systematic review on cellular responses of Escherichia coli to nonthermal electromagnetic irradiation
PublicationInvestigation of Escherichia coli under electromagnetic fields is of significance in human studies owing to its short doubling time and human‐like DNA mechanisms. The present review aims to systematically evaluate the literature to conclude causality between 0 and 300 GHz electromagnetic fields and biological effects in E. coli. To that end, the OHAT methodology and risk of bias tool were employed. Exponentially growing cells exposed...
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Molecular typing of clinical isolates of Escherichia coli by a new PCR MP method
PublicationTypowanie epidemiologiczne bakterii jest rutynową procedurą stosowaną w badaniach zakażeń szpitalnych i wymaga stosowania odpowiednich metod o wysokim potencjale różnicującym. Przedstawiliśmy możliwości nowej techniki PCR MP w różnicowaniu szczepów E. coli. Rezultaty zostały porównane z wynikami uzyskanymi metodą PFGE.
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Molecular typing of clinical isolates of Escherichia coli by a new PCR MP method
PublicationTypowanie epidemiologiczne bakterii jest rutynową procedurą stosowaną w badaniach zakażeń szpitalnych i wymaga stosowania odpowiednich metod o wysokim potencjale różnicującym. Przedstawiliśmy możliwości nowej techniki PCR MP w różnicowaniu szczepów E. coli. Rezultaty zostały porównane z wynikami uzyskanymi metodą PFGE.
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Leukemia and risk of recurrent Escherichia coli bacteremia: genotyping implicates E. coli translocation from the colon to the bloodstream.
PublicationIn patients with leukemia, the portal(s) and reasons for the persistence of an Escherichia coli recurrent bacteremia remain unclear. Adult Hematology Clinic (AHC) databases at the State Clinical Hospital in Gdańsk were reviewed to evaluate the frequency of E. coli bacteremia between 2002 and 2005. Blood and bowel E. coli strains were obtained and the genetic relatedness of the strains was analyzed. The rate of E. coli bacteremia...
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Regulated assembly of lipopolysaccharide and sensing of its alterations in Escherichia coli
PublicationThis thesis describes mechanism of the regulation of the transcription of the rpoE gene encoding an essential RNA polymerase subunit in Escherichia coli. The RpoE regulates extracytoplasmic stress response regulon and is required to initiate transcription of genes, whose products are involvedin the folding of periplasmic proteins and synthesis and transport of outer membrane components. The transcriptional regulation of the rpoE...
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Biofilm formation as a virulence determinant of uropathogenic Escherichia coli Dr+ strains
PublicationInfekcje dróg moczowych stanowią największy problem zdrowotny dotykający miliony osób populacji ludzkiej każdego roku. Uropatogenne szczepy Escherichia coli są najczęstszym czynnikiem etiologicznym odpowiedzialnym za wywoływanie górnych i dolnych dróg moczowych. Powyższe szczepy bakteryjne produkują szereg czynników wirulentnych pośród, których wyróżnić można eksponowane powierzchniowo polimeryczne struktury adhezyjne (fimbrie/pile)...
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Checkpoints that regulate balanced biosynthesis of lipopolysaccharide and its essentiality in Escherichia coli
PublicationThe outer membrane (OM) of Gram-negative bacteria, such as Escherichia coli, is essential for their viability. Lipopolysaccharide (LPS) constitutes the major component of OM, providing the permeability barrier, and a tight balance exists between LPS and phospholipids amounts as both of these essential components use a common metabolic precursor. Hence, checkpoints are in place, right from the regulation of the first committed step...
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Cloning of the thermostable alpha-amylase gene from Pyrococcus woesei in Escherichia coli.
PublicationGen kodujący alfa-amylazę z Pyrococcus woesei klonowano do plazmidów pET21d(+)lub pYTB2, którymi transformowano komórki Escherichia coli. Otrzymana rekombinantowa alfa amylaza wykazuje maksymalną aktywność przy pH 5.6 w temperaturze 95 stC i przejawia około 24 % początkowej aktywności nawet po 2 godz. inkubacji w 120 stC. Duża termostabilność otrzymanego preparatu świadczy o jego przydatności do enzymatycznego upłynniania skrobi.
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Resistance of Escherichia coli and Enterococcus spp. to selected antimicrobial agents present in municipal
PublicationIn this study, the susceptibility to erythromycin (E) and to trimethoprim/sulfamethoxazole (SXT) among isolates of Enterococcus spp. and Escherichia coli was tested, respectively. Both fecal indicators were detected and isolated from raw(RW) and treated wastewater (TW) as well as from samples of activated sludge (AS) collected in a local wastewater treatment plant (WWTP). Biodiversity of bacterial community in AS was also monitored...
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Isolation and some properties of the thermostable beta-galactosidase of Pyrococcus woesei expressed in Escherichia coli
PublicationOpracowano metodę izolowania oraz zbadano właściwości enzymu otrzymanego ze szczepu Escherichia coli zawierającego gen kodujący termostabilną beta-ga-laktozydazę z Pyrococcus woesei.
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Properties of recombinant trehalose synthase from Deinococcus radiodurans expressed in Escherichia coli
PublicationA 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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Use of Escherichia coli Nissle 1917 producing recombinant colicins for treatment of IBD patients
PublicationPatients 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...
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Cloning, Expression and Purification of Wild-Type Trehalose Synthase from Deinococcus geothermalis
PublicationThe aim of this study was isolation and cloning of trehalose synthase gene derived from extremophilic microorganism to the expression vectors in the Tabor-Studier system and its expression in Rosetta(DE3)pLysS Escherichia coli cells. The second phase of the study consisted of proteins purification using an initial denaturation of host proteins and salting-out proteins by ammonium sulfate.
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Assembly of Lipopolysaccharide in Escherichia coli Requires the Essential LapB Heat Shock Protein
PublicationHere, we describe two new heat shock proteins involved in the assembly of LPS in Escherichia coli, LapA and LapB (lipopolysaccharide assembly protein A and B). lapB mutants were identified based on an increased envelope stress response. Envelope stress responsive pathways control key steps in LPS biogenesis and respond to defects in the LPS assembly. Accordingly, the LPS content in ΔlapB or Δ(lapA lapB) mutants was elevated, with...
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Molecular aspects of biogenesis of Escherichia coli Dr Fimbriae: characterization of DraB-DraE complexes
PublicationHemaglutynina Dr uropatogennych szczepów Escherichia coli jest fimbrialnym homopolimerem zbudowanym z podjednostek białkowych DraE kodowanych przez operon dra. Operon dra zawiera geny draB i draC, kodujące białka homologiczne do białek (białko opiekuńcze-białko kanałotwórcze) zaangażowanych w procesie biogenezy struktur polimerycznych eksprymowanych powierzchniowo. Białko DraB jest jednym z białek periplazmatycznych, należącym...
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Alternative treatment approaches of urinary tract infections caused by uropathogenic Escherichia coli strains
PublicationUrinary tract infections (UTIs) are the most widespread and annoying infections affecting millions of people every year annually. The biggest problem of urinary diseases are recurrences, the increasing resistance of uropathogens to commonly used antibiotics, as well as the high health care costs of afflicted persons. Uropathogenic Escherichia coli strains (UPECs) are the most dominant etiologic agent of community-acquired infections...
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Characterization of immobilized Escherichia coli cells transformed by β-galactosidase gene from Pyrococcus woesei
PublicationThermostable β-galactosidase from Escherichia coli transformant containing theenzyme gene from Pyrococcus woesei was immobilized on alginate gel.The benefits of using whole bacterial cells not only exclude expensive, laborious proteinisolation and purification but also stabilize enzymes by cytosol components. Increase inproductivity of enzyme can be achieved by cells permeabilization. To increase permeability ofcytoplasmic membrane...
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Central carbon metabolism influences fidelity of DNA replication in Escherichia coli
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Molekularny mechanizm biogenezy fimbrii Dr uropatogennych szczepów Escherichia coli.
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Regulation of LPS assembly via controlled proteolysis and sensing of LPS stress in Escherichia coli
PublicationLipopolysaccharide (LPS) is a complex glycolipid, essential for the bacterial viability and along with phospholipids, it constitutes the major amphiphilic component of outer membrane (OM) in most of the Gram-negative bacteria, including Escherichia coli. LPS molecules confer an effective permeability barrier to the OM and play a crucial role in bacteria-environment and -host interactions. The synthesis and accumulation of this...
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Identification of Substrates of Cytoplasmic Peptidyl-Prolyl Cis/Trans Isomerases and Their Collective Essentiality in Escherichia Coli
PublicationProtein folding often requires molecular chaperones and folding catalysts, such as peptidyl-prolyl cis/trans isomerases (PPIs). The Escherichia coli cytoplasm contains six well-known PPIs, although a requirement of their PPIase activity, the identity of their substrates and relative enzymatic contribution is unknown. Thus, strains lacking all periplasmic and one of the cytoplasmic PPIs were constructed. Measurement of their PPIase...
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Isothiocyanates as effective agents against enterohemorrhagic Escherichia coli: insight to the mode of action
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Hydrophilic domain II of Escherichia coli Dr fimbriae facilitates cell invasion
PublicationStosując techniki mutagenezy ukierunkowanej oraz mikroskopii elektronowej stwierdzono, że hydrofilowa domena II białka podjednostkowego DraE fimbrii jest odpowiedzialna za inwazję komórki. Otrzymane wyniki są zgodne z interpretacją, że fenotypy fimbrii Dr wiązania i inwazji komórki HeLa mogą być oddzielone.
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Role of the disulfide bond in stabilizing and folding of the fimbrial protein DraE from uropathogenic Escherichia coli
PublicationDr fimbriae are homopolymeric adhesive organelles of uropathogenic Escherichia coli composed of DraE subunits, responsible for the attachment to host cells. These structures are characterized by enormously high stability resulting from the structural properties of an Ig-like fold of DraE. One feature of DraE and other fimbrial subunits that makes them peculiar among Ig-like domain-containing proteins is a conserved disulfide bond...