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Articoli di riviste sul tema "Escherichia coli Inclusions"

Autore: Grafiati

Pubblicato: 4 giugno 2021

Ultima modifica: 1 febbraio 2022

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1

Hänisch, Jan, Marc Wältermann, Horst Robenek e Alexander Steinbüchel. "The Ralstonia eutropha H16 phasin PhaP1 is targeted to intracellular triacylglycerol inclusions in Rhodococcus opacus PD630 and Mycobacterium smegmatis mc2155, and provides an anchor to target other proteins". Microbiology 152, n. 11 (1 novembre 2006): 3271–80. http://dx.doi.org/10.1099/mic.0.28969-0.

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In Ralstonia eutropha, the H16 phasin PhaP1 represents the major phasin that binds to the surface of polyhydroxyalkanoate (PHA) inclusions. In this study, C-terminal fusions of PhaP1 with enhanced green fluorescent protein (eGFP) and with Escherichia coli β-galactosidase (LacZ) were expressed separately in the triacylglycerol (TAG)-accumulating actinomycetes Rhodococcus opacus PD630 and Mycobacterium smegmatis mc2155, employing the M. smegmatis acetamidase (ace) promoter of the Escherichia–Mycobacterium/Rhodococcus shuttle plasmid pJAM2. PhaP1 and the PhaP1 fusion proteins were expressed stably in the recombinant strains. Western blot analysis of cell fractions of Rh. opacus revealed that PhaP1 and the PhaP1–eGFP fusion protein were associated with the TAG inclusions, whereas no phasin or phasin fusion protein was detected in the soluble and membrane fractions. Additional electron microscopy/immunocytochemistry studies demonstrated that PhaP1 was mainly located on the surface of intracellular TAG inclusions; in addition, some PhaP1 also occurred at the plasma membrane. Fluorescence microscopic investigations of the subcellular distribution of the PhaP1–eGFP fusion protein in vivo and on isolated TAG inclusions revealed that the fusion protein was bound to TAG inclusions at all stages of their formation, and to some extent at the cytoplasmic membrane. The PhaP1–LacZ fusion protein also bound to the TAG inclusions, and could be separated together with the inclusions from Rh. opacus crude extracts, thus demonstrating the immobilization of β-galactosidase activity on the inclusions. This is believed to be the first report demonstrating the ability of PhaP1 to bind to lipid inclusions in addition to PHA inclusions. Furthermore, it was demonstrated that this non-specificity of PhaP1 can be utilized to anchor enzymically active fusion proteins to a matrix of bacterial TAG inclusions.

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Chen, Shuxiong, Natalie A. Parlane, Jason Lee, D. Neil Wedlock, Bryce M. Buddle e Bernd H. A. Rehm. "New Skin Test for Detection of Bovine Tuberculosis on the Basis of Antigen-Displaying Polyester Inclusions Produced by Recombinant Escherichia coli". Applied and Environmental Microbiology 80, n. 8 (14 febbraio 2014): 2526–35. http://dx.doi.org/10.1128/aem.04168-13.

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ABSTRACTThe tuberculin skin test for diagnosing tuberculosis (TB) in cattle lacks specificity if animals are sensitized to environmental mycobacteria, as some antigens in purified protein derivative (PPD) prepared fromMycobacterium bovisare present in nonpathogenic mycobacteria. Three immunodominant TB antigens, ESAT6, CFP10, and Rv3615c, are present in members of the pathogenicMycobacterium tuberculosiscomplex but absent from the majority of environmental mycobacteria. These TB antigens have the potential to enhance skin test specificity. To increase their immunogenicity, these antigens were displayed on polyester beads by translationally fusing them to a polyhydroxyalkanoate (PHA) synthase which mediated formation of antigen-displaying inclusions in recombinantEscherichia coli. The most common form of these inclusions is poly(3-hydroxybutyric acid) (PHB). The respective fusion proteins displayed on these PHB inclusions (beads) were identified using tryptic peptide fingerprinting analysis in combination with matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS). The surface exposure and accessibility of antigens were assessed by enzyme-linked immunosorbent assay (ELISA). Polyester beads displaying all three TB antigens showed greater reactivity with TB antigen-specific antibody than did beads displaying only one TB antigen. This was neither due to cross-reactivity of antibodies with the other two antigens nor due to differences in protein expression levels between beads displaying single or three TB antigens. The triple-antigen-displaying polyester beads were used for skin testing of cattle and detected all cattle experimentally infected withM. boviswith no false-positive reactions observed in those sensitized to environmental mycobacteria. The results suggested applicability of TB antigen-displaying polyester inclusions as diagnostic reagents for distinguishing TB-infected from noninfected animals.

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Davis, Katelin L., Liang Cheng, José Ramos-Vara, Melissa D. Sánchez, Rebecca P. Wilkes e Mario F. Sola. "Malakoplakia in the Urinary Bladder of 4 Puppies". Veterinary Pathology 58, n. 4 (23 aprile 2021): 699–704. http://dx.doi.org/10.1177/03009858211009779.

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Malakoplakia in humans most often affects the urinary bladder and is characterized by inflammation with von Hansemann–type macrophages, with or without Michaelis-Gutmann bodies, and is frequently associated with Escherichia coli infection. We describe the microscopic features of malakoplakia in the urinary bladder of 4 puppies. In all cases, the lamina propria of the urinary bladder was markedly expanded by sheets of large, round to polygonal macrophages with intracytoplasmic, periodic acid-Schiff-positive granules and granular inclusions, and rare Prussian blue–positive inclusions. Macrophages were positive for CD18 and Iba1. In 2 cases, Michaelis-Gutmann bodies were detected with hematoxylin and eosin stain and were best demonstrated with von Kossa stain. E. coli infection was confirmed in 2 cases with bacterial culture or polymerase chain reaction (PCR) and sequencing of the bacterial 16S ribosomal RNA gene. Transmission electron microscopy of one case demonstrated macrophages with abundant lysosomes, phagolysosomes, and rod-shaped bacteria. Microscopic features were similar to human cases of malakoplakia. In dogs, the light microscopic characteristics of malakoplakia closely resemble granular cell tumors and histiocytic ulcerative colitis.

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Wada, Y., H. Kondo, Y. Nakaoka e M. Kubo. "Gastric Attaching and Effacing Escherichia coli Lesions in a Puppy with Naturally Occurring Enteric Colibacillosis and Concurrent Canine Distemper Virus Infection". Veterinary Pathology 33, n. 6 (novembre 1996): 717–20. http://dx.doi.org/10.1177/030098589603300615.

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A puppy suffering from chronic diarrhea was humanely killed at 90 days of age. Numerous Gram-negative bacilli were found adhering to the surface of as well as within epithelial cells from the stomach to the colon. Canine distemper virus inclusions were in the epithelial cytoplasm of the esophageal, gastric, and intestinal mucosa. Typical attaching and effacing ultrastructural lesions were in the stomach, and some bacilli were in the cytoplasm of the epithelial cells, Escherichia coli, isolated from the contents of the small intestine, belonged to serotype 0118: NM and were negative for plasmid-encoded EPEC adherence factor (EAF) and positive for the E. coli attaching effacing ( eae) gene. Immunohistologically, bacilli attached to the epithelium from the stomach to the colon were positive for antisera against E. coli 0118. E. coli0118: NM inoculated into human tissue culture cells (HEp-2 cells) were attached to the surface of the cells and within the cytoplasm. This is the first report of attaching and effacing E. coli (AEEC) infection in the stomach of the dog.

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Aldrich, H. C., S. Elvington, HE Machines, R. Szabady, K. Feder, L. McDowell e J. M. Shively. "Ultrastructural and Cytochemical Analyses of the Expression of the Thiobacillus Carboxysome Operon in Escherichia Coli". Microscopy and Microanalysis 7, S2 (agosto 2001): 740–41. http://dx.doi.org/10.1017/s1431927600029779.

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The cytoplasm of the bacterium Thiobacillus neapolitanus contains 117 nm diameter polyhedral inclusions, “carboxysomes” (Fig. 1) that contain ribulose-1,5- bisphosphate carboxylase/oxygenase (RuBisCO). Surrounding the polyhedron are nonmembranous proteinaceous plates devoid of lipid. The carboxysomes are composed of at least 8 major peptides, all coded within the same operon. Six (CsoSIA, CsoSIB, CsoSIC, CsoS2A, CsoS2B, and CsoS3) make up the shell, and two are the large (CbbL) and small subunits (CbbS) of RuBisCO. Using immunogold labeling on ultrathin sections, peptides CsoS2A, CsoS2B, and CsoS3 have been localized to the shell. Since the original characterization of the csoSl gene, we have also immunolocalized the CsoSl peptide to the shell.As part of our initial efforts to understand how these components are assembled into the symmetrical, functional entity, the carboxysome operon from T. neapolitanus was cloned into the pET-21a(+) plasmid, an expression vector that codes for resistance to ampicillin.

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Blatchford, Paul A., Colin Scott, Nigel French e Bernd H. A. Rehm. "Immobilization of organophosphohydrolase OpdA from Agrobacterium radiobacter by overproduction at the surface of polyester inclusions inside engineered Escherichia coli". Biotechnology and Bioengineering 109, n. 5 (26 dicembre 2011): 1101–8. http://dx.doi.org/10.1002/bit.24402.

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Kalscheuer, Rainer, Tim Stöveken, Heinrich Luftmann, Ursula Malkus, Rudolf Reichelt e Alexander Steinbüchel. "Neutral Lipid Biosynthesis in Engineered Escherichia coli: Jojoba Oil-Like Wax Esters and Fatty Acid Butyl Esters". Applied and Environmental Microbiology 72, n. 2 (febbraio 2006): 1373–79. http://dx.doi.org/10.1128/aem.72.2.1373-1379.2006.

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ABSTRACT Wax esters are esters of long-chain fatty acids and long-chain fatty alcohols which are of considerable commercial importance and are produced on a scale of 3 million tons per year. The oil from the jojoba plant (Simmondsia chinensis) is the main biological source of wax esters. Although it has a multitude of potential applications, the use of jojoba oil is restricted, due to its high price. In this study, we describe the establishment of heterologous wax ester biosynthesis in a recombinant Escherichia coli strain by coexpression of a fatty alcohol-producing bifunctional acyl-coenzyme A reductase from the jojoba plant and a bacterial wax ester synthase from Acinetobacter baylyi strain ADP1, catalyzing the esterification of fatty alcohols and coenzyme A thioesters of fatty acids. In the presence of oleate, jojoba oil-like wax esters such as palmityl oleate, palmityl palmitoleate, and oleyl oleate were produced, amounting to up to ca. 1% of the cellular dry weight. In addition to wax esters, fatty acid butyl esters were unexpectedly observed in the presence of oleate. The latter could be attributed to solvent residues of 1-butanol present in the medium component, Bacto tryptone. Neutral lipids produced in recombinant E. coli were accumulated as intracytoplasmic inclusions, demonstrating that the formation and structural integrity of bacterial lipid bodies do not require specific structural proteins. This is the first report on substantial biosynthesis and accumulation of neutral lipids in E. coli, which might open new perspectives for the biotechnological production of cheap jojoba oil equivalents from inexpensive resources employing recombinant microorganisms.

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Petrus, Marloes L. C., Lukas A. Kiefer, Pranav Puri, Evert Heemskerk, Michael S. Seaman, Dan H. Barouch, Sagrario Arias, Gilles P. van Wezel e Menzo Havenga. "A microbial expression system for high-level production of scFv HIV-neutralizing antibody fragments in Escherichia coli". Applied Microbiology and Biotechnology 103, n. 21-22 (22 ottobre 2019): 8875–88. http://dx.doi.org/10.1007/s00253-019-10145-1.

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Abstract Monoclonal antibodies (mABs) are of great biopharmaceutical importance for the diagnosis and treatment of diseases. However, their production in mammalian expression hosts usually requires extensive production times and is expensive. Escherichia coli has become a new platform for production of functional small antibody fragment variants. In this study, we have used a rhamnose-inducible expression system that allows precise control of protein expression levels. The system was first evaluated for the cytoplasmic production of super folder green fluorescence protein (sfGFP) in various production platforms and then for the periplasmic production of the anti-HIV single-chain variable antibody fragment (scFv) of PGT135. Anti-HIV broadly neutralizing antibodies, like PGT135, have potential for clinical use to prevent HIV transmission, to promote immune responses and to eradicate infected cells. Different concentrations of L-rhamnose resulted in the controlled production of both sfGFP and scFv PGT135 antibody. In addition, by optimizing the culture conditions, the amount of scFv PGT135 antibody that was expressed soluble or as inclusions bodies could be modulated. The proteins were produced in batch bioreactors, with yields of 4.9 g/L for sfGFP and 0.8 g/L for scFv. The functionality of the purified antibodies was demonstrated by their ability to neutralize a panel of different HIV variants in vitro. We expect that this expression system will prove very useful for the development of a more cost-effective production process for proteins and antibody fragments in microbial cells.

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Carija, Pinheiro, Iglesias e Ventura. "Computational Assessment of Bacterial Protein Structures Indicates a Selection Against Aggregation". Cells 8, n. 8 (8 agosto 2019): 856. http://dx.doi.org/10.3390/cells8080856.

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The aggregation of proteins compromises cell fitness, either because it titrates functional proteins into non-productive inclusions or because it results in the formation of toxic assemblies. Accordingly, computational proteome-wide analyses suggest that prevention of aggregation upon misfolding plays a key role in sequence evolution. Most proteins spend their lifetimes in a folded state; therefore, it is conceivable that, in addition to sequences, protein structures would have also evolved to minimize the risk of aggregation in their natural environments. By exploiting the AGGRESCAN3D structure-based approach to predict the aggregation propensity of >600 Escherichia coli proteins, we show that the structural aggregation propensity of globular proteins is connected with their abundance, length, essentiality, subcellular location and quaternary structure. These data suggest that the avoidance of protein aggregation has contributed to shape the structural properties of proteins in bacterial cells.

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Rybalchenko, O. V., O. G. Orlova, L. B. Zakharova, O. N. Vishnevskaya e A. G. Markov. "EFFECT OF PROBIOTIC BACTERIA AND LIPOPOLISACCHARIDES ON EPITELIOCYTES TIGHT JUNCTIONS OF RAT JEJUNUM". Journal of microbiology epidemiology immunobiology, n. 6 (28 dicembre 2017): 80–87. http://dx.doi.org/10.36233/0372-9311-2017-6-80-87.

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Aim. The present study has been undertaken with the main objective the influence of probiotic bacteria Lactobacillus plantarum 8 РАЗ and Escherichia coli M17 and lipopolysaccharide on the ultrastructure of enterocytes tight junctions of mucous membranes of rat jejunum. Materials and methods. The study was carried out on E. coli lipopolysaccharide (Sigma-Aldrich, Germany) and probiotic bacteria L. plantarum 8PA3 and E. coli M17. Male Wistar rats were used. A comparative analysis of the ultrathin structure of enterocytes and tight junctions were carried out by successive incubation of rat jejunum with probiotic bacteria L. plantarum 8PA3 and E. coli Ml 7, with lipopolysaccharide and a complex of bacteria with LPS. Results. The effect of L. plantarum 8PA3 on the mucosa of rats jejunum on a number of characters was similar to E. coli Mil. It manifested by preservation of the intact structure of the intercellular space and tight junctions. At the same time, hollow spherical inclusions with fragments of bacteria surrounded by membranes detected in the cytoplasm of enterocytes testified to the possibility of penetration of probiotic bacteria through the mucous membrane of the jejunum by a transcellular pathway With simultaneous action on enterocytes of rats jejunum of probiotic bacteria and lipopolysaccharide complex no destructive changes in the structure of dense contacts were observed, however, in a significant number of cases, bacterial cells were found in the intercellular space next to the goblet cells. Conclusion. A similar effect of Gram-positive bacteria L. plantarum 8PA3 and Gram-negative bacteria E. coli Ml 7 and their complexes with lipopolysaccharide on the jejunum epitheliocytes was revealed. Morphological analysis showed that lipopolysaccharide might influence on parasel-lular transport by probiotic bacteria. In the absence of LPS, probiotic bacteria can possibly penetrate the mucosa of rats jejunum by a transcellular pathway.

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Kushnir, I. M., G. V. Kolodiy, V. I. Kushnir, S. D. Murska, I. S. Semen e U. Z. Berbeka. "THE INFLUENCE OF POLYHEXAMETHYLENE GUANIDINE SALTS ON THE MICROBIOLOGICAL PARAMETERS OF WATER". Scientific and Technical Bulletin оf State Scientific Research Control Institute of Veterinary Medical Products and Fodder Additives аnd Institute of Animal Biology 22, n. 1 (29 marzo 2021): 126–30. http://dx.doi.org/10.36359/scivp.2021-22-1.14.

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The article represent the use of disinfectant which was made based on salts of polyhexamethylene guanidine hydrochloride (PGMG-hydrochloride) and polyhexamethylene guanidine phosphate (PGMG-phosphate) for water disinfection. Water plays one of the most important roles in almost all life processes. Water is a part of the blood, participates in blood circulatory system, delivers nutrients and oxygen to the organs, participates in oxidation reaction, hydrolysis and other reactions of intercellular metabolism, forms an environment to maintain a healthy microflora of the digestive tract, which provides effective breakdown of nutrients by the optimal amount of enzymes. Good drinking water quality must to be epidemically safe and free of pathogenic microbes, viruses and other biological inclusions. Also must be harmless in chemical composition, have good organoleptic properties ‒ be transparent, colorless, have no taste or odor. Water quality is assessed by total microbial contamination and the amount of bacteria of the Escherichia coli group in 1 cm3 of water, and also by thermostable Escherichia coli (fecal coliforms) in 100 cm3 of water, pathogenic microorganisms and the amount of coli-phages. In result of the conducted researches it was found that the use of disinfectant in 0,2 % concentration did not reduce the total microbial pollution of water. In particular, the efficiency of 1, 2 and 3-hour of exposure was 57, 62 and 68 %, respectively. After application of disinfectant in 0,3 % concentration the efficiency of its application was for 1 h – 69 %, 2 h – 82 %, 3 h – 100 %. It was found that the use of a disinfectant, which was made based on PGMG salts in 0,4 % concentration contributed to reduce the total microbial contamination of water. The efficiency by the 1, 2 and 3 hour of exposure was, respectively, 73, 86 and 100 %. Therefore, the use of a disinfectant, containing in its composition PGMG salts, in 0,3% concentration at 3 hours of exposure was more optimal, which contributed to reduce the total microbial contamination of water.

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Park, Youngjin, Mohd Amir F. Abdullah, Milton D. Taylor, Khalidur Rahman e Michael J. Adang. "Enhancement of Bacillus thuringiensis Cry3Aa and Cry3Bb Toxicities to Coleopteran Larvae by a Toxin-Binding Fragment of an Insect Cadherin". Applied and Environmental Microbiology 75, n. 10 (27 marzo 2009): 3086–92. http://dx.doi.org/10.1128/aem.00268-09.

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ABSTRACT The Cry3Aa and Cry3Bb insecticidal proteins of Bacillus thuringiensis are used in biopesticides and transgenic crops to control larvae of leaf-feeding beetles and rootworms. Cadherins localized in the midgut epithelium are identified as receptors for Cry toxins in lepidopteran and dipteran larvae. Previously, we discovered that a peptide of a toxin-binding cadherin expressed in Escherichia coli functions as a synergist for Cry1A toxicity against lepidopteran larvae and Cry4 toxicity against dipteran larvae. Here we report that the fragment containing the three most C-terminal cadherin repeats (CR) from the cadherin of the western corn rootworm binds toxin and enhances Cry3 toxicity to larvae of naturally susceptible species. The cadherin fragment (CR8 to CR10 [CR8-10]) of western corn rootworm Diabrotica virgifera virgifera was expressed in E. coli as an inclusion body. By an enzyme-linked immunosorbent microplate assay, we demonstrated that the CR8-10 peptide binds α-chymotrypsin-treated Cry3Aa and Cry3Bb toxins at high affinity (11.8 nM and 1.4 nM, respectively). Coleopteran larvae ingesting CR8-10 inclusions had increased susceptibility to Cry3Aa or Cry3Bb toxin. The Cry3 toxin-enhancing effect of CR8-10 was demonstrated for Colorado potato beetle Leptinotarsa decemlineata, southern corn rootworm Diabrotica undecimpunctata howardi, and western corn rootworm. The extent of Cry3 toxin enhancement, which ranged from 3- to 13-fold, may have practical applications for insect control. Cry3-containing biopesticides that include a cadherin fragment could be more efficacious. And Bt corn (i.e., corn treated with B. thuringiensis to make it resistant to pests) coexpressing Cry3Bb and CR8-10 could increase the functional dose level of the insect toxic activity, reducing the overall resistance risk.

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Tam, Jeffrey E., Carolyn H. Davis e Priscilla B. Wyrick. "Expression of recombinant DNA introduced into Chlamydia trachomatis by electroporation". Canadian Journal of Microbiology 40, n. 7 (1 luglio 1994): 583–91. http://dx.doi.org/10.1139/m94-093.

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Electroporation was used to introduce DNA into the elementary bodies of the obligate parasitic bacterium Chlamydia trachomatis. The source of DNA for these experiments was the chimeric plasmid pPBW100, which was constructed from the well-characterized 7.5-kb plasmid of C. trachomatis and the Escherichia coli plasmid pBGS9. To select directly for C. trachomatis carrying pPBW100, an in-frame gene fusion between the chlamydial promoter P7248 and a promoterless chloramphenicol acetyltransferase (cat) cassette was incorporated into the plasmid. After infection of McCoy cells with electroporated elementary bodies containing pPBW100, the following were observed: (i) the plasmid DNA was detected inside the chloramphenicol-resistant chlamydial inclusions by in situ and Southern hybridization analyses; (ii) both physical and biochemical evidence showed that chloramphenicol acetyltransferase was synthesized by the electroporated C. trachomatis; (iii) expression of P7248::cat was developmentally regulated and occurred during the early stages of chlamydial reticulate body development; and (iv) although the expression from P7248::cat was mainly transient, there were rare instances where chloramphenicol-resistant C. trachomatis were observed after four passages.Key words: chlamydia, electroporation, chimeric plasmid, expression.

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Mifune, Jun, Katrin Grage e Bernd H. A. Rehm. "Production of Functionalized Biopolyester Granules by Recombinant Lactococcus lactis". Applied and Environmental Microbiology 75, n. 14 (22 maggio 2009): 4668–75. http://dx.doi.org/10.1128/aem.00487-09.

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ABSTRACT Many bacteria are naturally capable of accumulating biopolyesters composed of 3-hydroxy fatty acids as intracellular inclusions, which serve as storage granules. Recently, these inclusions have been considered as nano-/microbeads with surface-attached proteins, which can be engineered to display various protein-based functions that are suitable for biotechnological and biomedical applications. In this study, the food-grade, generally-regarded-as-safe gram-positive organism Lactococcus lactis was engineered to recombinantly produce the biopolyester poly(3-hydroxybutyrate) and the respective intracellular inclusions. The codon-optimized polyhydroxybutyrate biosynthesis operon phaCAB from Cupriavidus necator was expressed using the nisin-controlled gene expression system. Recombinant L. lactis accumulated up to 6% (wt/wt) poly(3-hydroxybutyrate) of cellular dry weight. Poly(3-hydroxybutyrate) granules were isolated and analyzed with respect to bound proteins using biochemical methods and with respect to shape/size using transmission electron microscopy. The immunoglobulin G (IgG) binding ZZ domain of Staphylococcus aureus protein A was chosen as an exemplary functionality to be displayed at the granule surface by fusing it to the N terminus of the granule-associated poly(3-hydroxybutyrate) synthase. The presence of the fusion protein at the surface of isolated granules was confirmed by peptide fingerprinting using matrix-assisted laser desorption ionization-time of flight (mass spectrometry). The functionality of the ZZ domain-displaying granules was demonstrated by enzyme-linked immunosorbent assay and IgG affinity purification. In both assays, the ZZ beads from recombinant L. lactis performed at least equally to ZZ beads from Escherichia coli. Overall, in this study it was shown that recombinant L. lactis can be used to manufacture endotoxin-free poly(3-hydroxybutyrate) beads with surface functionalities that are suitable for biomedical applications.

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Parlane, Natalie A., D. Neil Wedlock, Bryce M. Buddle e Bernd H. A. Rehm. "Bacterial Polyester Inclusions Engineered To Display Vaccine Candidate Antigens for Use as a Novel Class of Safe and Efficient Vaccine Delivery Agents". Applied and Environmental Microbiology 75, n. 24 (16 ottobre 2009): 7739–44. http://dx.doi.org/10.1128/aem.01965-09.

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ABSTRACT Bioengineered bacterial polyester inclusions have the potential to be used as a vaccine delivery system. The biopolyester beads were engineered to display a fusion protein of the polyester synthase PhaC and the two key antigens involved in immune response to the infectious agent that causes tuberculosis, Mycobacterium tuberculosis, notably antigen 85A (Ag85A) and the 6-kDa early secreted antigenic target (ESAT-6) from Mycobacterium tuberculosis. Polyester beads displaying the respective fusion protein at a high density were successfully produced (henceforth called Ag85A-ESAT-6 beads) by recombinant Escherichia coli. The ability of the Ag85A-ESAT-6 beads to enhance mouse immunity to the displayed antigens was investigated. The beads were not toxic to the animals, as determined by weight gain and absence of lesions at the inoculation site in immunized animals. In vivo injection of the Ag85A-ESAT-6 beads in mice induced significant humoral and cell-mediated immune responses to both Ag85A and ESAT-6. Vaccination with Ag85A-ESAT-6 beads was efficient at stimulating immunity on their own, and this ability was enhanced by administration of the beads in an oil-in-water emulsion. In addition, vaccination with the Ag85A-ESAT-6 beads induced significantly stronger humoral and cell-mediated immune responses than vaccination with an equivalent dose of the fusion protein Ag85A-ESAT-6 alone. The immune response induced by the beads was of a mixed Th1/Th2 nature, as assessed from the induction of the cytokine gamma interferon (Th1 immune response) and increased levels of immunoglobulin G1 (Th2 immune response). Hence, engineered biopolyester beads displaying foreign antigens represent a new class of versatile, safe, and biocompatible vaccines.

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Kim, Won-Seok, Jeong Sun-Hyung, Ro-Dong Park, Kil-Yong Kim e Hari B. Krishnan. "Sinorhizobium fredii USDA257 Releases a 22-kDa Outer Membrane Protein (Omp22) to the Extracellular Milieu When Grown in Calcium-Limiting Conditions". Molecular Plant-Microbe Interactions® 18, n. 8 (agosto 2005): 808–18. http://dx.doi.org/10.1094/mpmi-18-0808.

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Calcium, which regulates a wide variety of cellular functions, plays an important role in Rhizobium-legume interactions. We investigated the effect of calcium on surface appendages of Sinorhizobium fredii USDA257. Cold-field emission scanning electron microscopy observation of USDA257 grown in calcium-limiting conditions revealed cells with unusual shape and size. Transmission electron microscopy observation revealed intact flagella were present only when USDA257 cells were grown in calcium-sufficient conditions. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of flagellar preparations from USDA257 cells grown in calcium-limiting conditions showed the presence of a 22-kDa protein that was absent from cells grown in calcium-sufficient conditions. We have cloned and determined the nucleotide sequence of the gene encoding the 22- kDa protein. After successful expression in Escherichia coli, polyclonal antibodies were raised against the recombinant 22-kDa protein (Omp22). Subcellular fractionation analysis demonstrated that Omp22 was predominantly present in the extracellular fraction. Western blot analysis revealed the presence of immunologically related proteins from diverse rhizobia. Immunocytochemical localization of thin sections of USDA257 cells showed specific labeling of protein A-gold particles on protein inclusions found proximal to the cells. Accumulation of Omp22 was greatly reduced when USDA257 cells were grown in the presence of increasing calcium. Northern blot analysis indicated that calcium was the only divalent cation among those tested that down-regulated omp22 expression. An omp22 mutant was able to grow in calcium-limiting conditions at a rate similar to that of wild-type USDA257. Significantly more nodules were initiated by the omp22 mutant than by the wild-type on soybean cultivar Peking grown in calciumlimiting conditions.

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Gorbatuk, O. B., U. S. Nikolayev, D. M. Irodov, I. Ya Dubey e P. V. Gilchuk. "Refolding of ScFv-CBD fusion protein from Escherichia coli inclusion bodies". Biopolymers and Cell 24, n. 1 (20 gennaio 2008): 51–59. http://dx.doi.org/10.7124/bc.000790.

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Steinmann, Björn, Andreas Christmann, Tim Heiseler, Janine Fritz e Harald Kolmar. "In Vivo Enzyme Immobilization by Inclusion Body Display". Applied and Environmental Microbiology 76, n. 16 (25 giugno 2010): 5563–69. http://dx.doi.org/10.1128/aem.00612-10.

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ABSTRACT A novel strategy for in vivo immobilization of enzymes on the surfaces of inclusion bodies has been established. It relies on expression in Escherichia coli of the polyhydroxybutyrate synthase PhaC from Cupriavidus necator, which carries at its amino terminus an engineered negatively charged α-helical coil (Ecoil) and forms inclusion bodies upon high-level expression. Coexpression in the same cell of galactose oxidase (GOase) from Fusarium spp. carrying a carboxy-terminal positively charged coil (lysine-rich coil [Kcoil]) sequence results in heterodimeric coiled-coil formation in vivo and in the capture of the enzyme in active form on the surface of the inclusion body particle. These round-shaped enzyme-decorated microparticles, with sizes of approximately 0.7 μm, can be isolated from lysed cells simply by centrifugation. The cost-effective one-step generation and isolation of enzymes immobilized on inclusion body particles may become useful for various applications in bioprocessing and biotransformation.

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19

Johnson, Dustin L., Chris B. Stone e James B. Mahony. "Interactions between CdsD, CdsQ, and CdsL, Three Putative Chlamydophila pneumoniae Type III Secretion Proteins". Journal of Bacteriology 190, n. 8 (15 febbraio 2008): 2972–80. http://dx.doi.org/10.1128/jb.01997-07.

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ABSTRACT Chlamydophila pneumoniae is a gram-negative obligate intracellular bacterial pathogen that causes pneumonia and bronchitis and may contribute to atherosclerosis. The developmental cycle of C. pneumoniae includes a morphological transition from an infectious extracellular elementary body (EB) to a noninfectious intracellular reticulate body (RB) that divides by binary fission. The C. pneumoniae genome encodes a type III secretion (T3S) apparatus that may be used to infect eukaryotic cells and to evade the host immune response. In the present study, Cpn0712 (CdsD), Cpn0704 (CdsQ), and Cpn0826 (CdsL), three C. pneumoniae genes encoding yersiniae T3S YscD, YscQ, and YscL homologs, respectively, were cloned and expressed as histidine- and glutathione S-transferase (GST)-tagged proteins in Escherichia coli. Purified recombinant proteins were used to raise hyper-immune polyclonal antiserum and were used in GST pull-down and copurification assays to identify protein-protein interactions. CdsD was detected in both EB and RB lysates by Western blot analyses, and immunofluorescent staining demonstrated the presence of CdsD within inclusions. Triton X-114 solubilization and phase separation of chlamydial EB proteins indicated that CdsD partitions with cytoplasmic proteins, suggesting it is not an integral membrane protein. GST pull-down assays indicated that recombinant CdsD interacts with CdsQ and CdsL, and copurification assays with chlamydial lysates confirmed that native CdsD interacts with CdsQ and CdsL. To the best of our knowledge, this is the first report demonstrating interactions between YscD, YscQ, and YscL homologs of bacterial T3S systems. These novel protein interactions may play important roles in the assembly or function of the chlamydial T3S apparatus.

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20

Tzeng, Yih-Ling, Anup K. Datta, Cristy A. Strole, Michael A. Lobritz, Russell W. Carlson e David S. Stephens. "Translocation and Surface Expression of Lipidated Serogroup B Capsular Polysaccharide in Neisseria meningitidis". Infection and Immunity 73, n. 3 (marzo 2005): 1491–505. http://dx.doi.org/10.1128/iai.73.3.1491-1505.2005.

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ABSTRACT The capsule of N. meningitidis serogroup B, (α2→8)-linked polysialic acid and the capsules of other meningococcal serogroups and of other gram-negative bacterial pathogens are anchored in the outer membrane through a 1,2-diacylglycerol moiety. Previous work on the meningococcal cps complex in Escherichia coli K-12 indicated that deletion of genes designated lipA and lipB caused intracellular accumulation of hyperelongated capsule polymers lacking the phospholipid substitution. To better understand the role of lip and lipB in capsule expression in a meningococcal background, the location, sequence, and relationship to related bacterial capsule genes were defined and specific mutations in lipA and lipB were generated in the serogroup B meningococcal strain NMB. The lipA and lipB genes are located on the 3′ end of the ctr operon and are most likely transcribed independently. Inactivation of lipA, lipB, and both resulted in the same total levels of capsular polymer production as in the parental controls; however, these mutants were as sensitive as an unencapsulated mutant to killing by normal human serum. Immunogold electron microscopy and flow cytometric analyses revealed intracellular inclusions of capsular polymers in lipA, lipB, and lipA lipB mutants. Capsular polymers purified from lipA, lipB, and lipA lipB mutants were lipidated. The phospholipid anchor was shown by gas chromatography-mass spectroscopy analysis to be a phosphodiester-linked 1,2-dipalmitoyl (C16:0) glycerol moiety and was identical in structure to that found on the wild-type meningococcal capsule polymers. Thus, lipA and lipB do not encode proteins responsible for diacylglycerophosphatidic acid substitution of the meningococcal capsule polymer; rather, they are required for proper translocation and surface expression of the lipidated polymer.

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21

Маркелова, Н. Ю., e N. Yu Markelova. "REP-elements of the Escherichia coli Genome and Transcription Signals: Positional and Functional Analysis". Mathematical Biology and Bioinformatics 10, n. 1 (24 giugno 2015): 245–59. http://dx.doi.org/10.17537/2015.10.245.

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In the intergenic regions of the Escherichia coli genome there are 356 REP- elements, containing 1-12 repeated sequences with degenerated consensus. Their biological role is poorly understood, but multiplicity in the genome, preferential localization between convergent genes and ability to form hairpin structures have led to the assumption that REP-elements participate in the transcription termination and processes affecting stability of the corresponding RNAs. Though the direct experiments did not confirm the ability of the model REP-sequence to stop RNA synthesis leaving some ambiguity regarding their primary function. In this study, positional and functional analysis was undertaken for the entire set of annotated REP-sequences and the reduced efficiency of RNA synthesis behind the many REP-modules was observed. However, some REP-modules had no effect on the transcription processivity, assuming the inclusion of REP-sequences into RNA and the possibility of their regulatory action. We also observed REP-associated transcription activation and found overlapping promoters. The most unexpected was specific distribution of REP-sequences nearby promoter islands, which indicates their insulator-like action maintaining functional autonomy of the “islands” and assumes functional significance of “island”-born RNAs.

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22

Jürgen, Britta, Antje Breitenstein, Vlada Urlacher, Knut Büttner, Hongying Lin, Michael Hecker, Thomas Schweder e Peter Neubauer. "Quality control of inclusion bodies in Escherichia coli". Microbial Cell Factories 9, n. 1 (2010): 41. http://dx.doi.org/10.1186/1475-2859-9-41.

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23

HARTLEY, D. L., e J. F. KANE. "Properties of inclusion bodies from recombinant Escherichia coli". Biochemical Society Transactions 16, n. 2 (1 aprile 1988): 101–2. http://dx.doi.org/10.1042/bst0160101.

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24

Gilchuk, P. V. "Evaluation of renaturation methods for industrial obtaining of recombinant proteins from Escherichia coli inclusion bodies in biologically active form". Biopolymers and Cell 20, n. 3 (20 maggio 2004): 182–92. http://dx.doi.org/10.7124/bc.0006a5.

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25

Simpson, R. J. "Solubilization of Escherichia coli Recombinant Proteins from Inclusion Bodies". Cold Spring Harbor Protocols 2010, n. 9 (1 settembre 2010): pdb.prot5485. http://dx.doi.org/10.1101/pdb.prot5485.

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26

Kane, James F., e Donna L. Hartley. "Formation of recombinant protein inclusion bodies in Escherichia coli". Trends in Biotechnology 6, n. 5 (maggio 1988): 95–101. http://dx.doi.org/10.1016/0167-7799(88)90065-0.

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27

Upadhyay, Vaibhav, Anupam Singh e Amulya K. Panda. "Purification of recombinant ovalbumin from inclusion bodies of Escherichia coli". Protein Expression and Purification 117 (gennaio 2016): 52–58. http://dx.doi.org/10.1016/j.pep.2015.09.015.

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28

Bowden, Gregory A., Angel M. Paredes e George Georgiou. "Structure and Morphology of Protein Inclusion Bodies in Escherichia Coli". Nature Biotechnology 9, n. 8 (agosto 1991): 725–30. http://dx.doi.org/10.1038/nbt0891-725.

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29

Rueda, Fabián, Olivia Cano-Garrido, Uwe Mamat, Kathleen Wilke, Joaquin Seras-Franzoso, Elena García-Fruitós e Antonio Villaverde. "Production of functional inclusion bodies in endotoxin-free Escherichia coli". Applied Microbiology and Biotechnology 98, n. 22 (17 agosto 2014): 9229–38. http://dx.doi.org/10.1007/s00253-014-6008-9.

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30

Carrió, M. Mar, e Antonio Villaverde. "Localization of Chaperones DnaK and GroEL in Bacterial Inclusion Bodies". Journal of Bacteriology 187, n. 10 (15 maggio 2005): 3599–601. http://dx.doi.org/10.1128/jb.187.10.3599-3601.2005.

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ABSTRACT By immunostaining and transmission electron microscopy, chaperones DnaK and GroEL have been identified at the solvent-exposed surface of bacterial inclusion bodies and entrapped within these aggregates, respectively. Functional implications of this distinct localization are discussed in the context of Escherichia coli protein quality control.

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31

Allam, Ayman B., Leticia Reyes, Nacyra Assad-Garcia, John I. Glass e Mary B. Brown. "Enhancement of Targeted Homologous Recombination in Mycoplasma mycoides subsp. capri by Inclusion of Heterologous recA". Applied and Environmental Microbiology 76, n. 20 (27 agosto 2010): 6951–54. http://dx.doi.org/10.1128/aem.00056-10.

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ABSTRACT A suicide plasmid, pExp1-ctpA::tetM-recAec, employing recA from Escherichia coli and tetM as a selection marker, was used to generate ctpA knockout mutants in Mycoplasma mycoides subsp. capri through targeted gene disruption. Inclusion of E. coli recA greatly enhanced both the consistency and the recovery of mutants generated by homologous recombination.

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32

Hart, R. A., U. Rinas e J. E. Bailey. "Protein composition of Vitreoscilla hemoglobin inclusion bodies produced in Escherichia coli." Journal of Biological Chemistry 265, n. 21 (luglio 1990): 12728–33. http://dx.doi.org/10.1016/s0021-9258(19)38405-4.

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33

McCaman, M. T. "Fragments of prochymosin produced in Escherichia coli form insoluble inclusion bodies." Journal of Bacteriology 171, n. 2 (1989): 1225–27. http://dx.doi.org/10.1128/jb.171.2.1225-1227.1989.

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34

Carretas-Valdez, Manuel I., Francisco J. Cinco-Moroyoqui, Marina J. Ezquerra-Brauer, Enrique Marquez-Rios, Idania E. Quintero-Reyes, Alonso A. Lopez-Zavala e Aldo A. Arvizu-Flores. "Refolding and Activation from Bacterial Inclusion Bodies of Trypsin I from Sardine (Sardinops sagax caerulea)". Protein & Peptide Letters 26, n. 3 (15 marzo 2019): 170–75. http://dx.doi.org/10.2174/0929866525666181019161114.

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Background: Trypsin from fish species is considered as a cold-adapted enzyme that may find potential biotechnological applications. In this work, the recombinant expression, refolding and activation of Trypsin I (TryI) from Monterey sardine (Sardinops sagax caerulea) are reported. Methods: TryI was overexpressed in Escherichia coli BL21 as a fusion protein of trypsinogen with thioredoxin. Refolding of trypsinogen I was achieved by dialysis of bacterial inclusion bodies with a recovery of 16.32 mg per liter of Luria broth medium. Results: Before activation, the trypsinogen fusion protein did not show trypsin activity. Trypsinogen I was activated by adding 0.002 U of native TryI purified from the sardine pyloric caeca (nonrecombinant). The activated recombinant trypsin showed three times more activity than the nonrecombinant trypsin alone. Conclusion: The described protocol allowed obtaining sufficient amounts of recombinant TryI from Monterey sardine fish for further biochemical and biophysical characterization of its coldadaptation parameters.

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35

Di Lorenzo, Mirella, Aurelio Hidalgo, Michael Haas e Uwe T. Bornscheuer. "Heterologous Production of Functional Forms of Rhizopus oryzae Lipase in Escherichia coli". Applied and Environmental Microbiology 71, n. 12 (dicembre 2005): 8974–77. http://dx.doi.org/10.1128/aem.71.12.8974-8977.2005.

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ABSTRACT To date, expression of the lipase from Rhizopus oryzae (ROL) in Escherichia coli always led to the formation of inclusion bodies and inactive protein. However, the production of active ROL and its precursor ProROL in soluble form was achieved when E. coli Origami(DE3) and pET-11d were used as expression systems.

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36

Lee, Sang-Eun. "Galactooligosaccharide Synthesis by Active β-Galactosidase Inclusion Bodies-Containing Escherichia coli Cells". Journal of Microbiology and Biotechnology 21, n. 11 (28 novembre 2011): 1151–58. http://dx.doi.org/10.4014/jmb.1105.05021.

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37

Morreale, G. "Continuous processing of fusion protein expressed as an Escherichia coli inclusion body". Journal of Chromatography B 786, n. 1-2 (25 marzo 2003): 237–46. http://dx.doi.org/10.1016/s1570-0232(02)00718-3.

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38

Lipničanová, Sabina, Daniela Chmelová, Andrej Godány, Miroslav Ondrejovič e Stanislav Miertuš. "Purification of viral neuraminidase from inclusion bodies produced by recombinant Escherichia coli". Journal of Biotechnology 316 (giugno 2020): 27–34. http://dx.doi.org/10.1016/j.jbiotec.2020.04.005.

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39

Hwang, Soon Ook. "Effect of inclusion bodies on the buoyant density of recombinant Escherichia coli". Biotechnology Techniques 10, n. 3 (marzo 1996): 157–60. http://dx.doi.org/10.1007/bf00158938.

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40

Ni, He, Peng-Cheng Guo, Wei-Ling Jiang, Xiao-Min Fan, Xiang-Yu Luo e Hai-Hang Li. "Expression of nattokinase in Escherichia coli and renaturation of its inclusion body". Journal of Biotechnology 231 (agosto 2016): 65–71. http://dx.doi.org/10.1016/j.jbiotec.2016.05.034.

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41

Xia, Xiao-Xia, Ya-Ling Shen e Dong-Zhi Wei. "Purification and Characterization of Recombinant sTRAIL Expressed in Escherichia coli". Acta Biochimica et Biophysica Sinica 36, n. 2 (1 febbraio 2004): 118–22. http://dx.doi.org/10.1093/abbs/36.2.118.

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Abstract As a potential anti-tumor protein, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has drawn considerable attention. This report presented the purification and characterization of soluble TRAIL, expressed as inclusion bodies in E. coli. sTRAIL inclusion bodies were solubilized and refolded at a high concentration up to 0.9 g/L by a simple dilution method. Refolded protein was purified to electrophoretic homogeneity by a single-step immobilized metal affinity chromatography. The purified sTRAIL had a strong cytotoxic activity against human pancreatic tumor cell line 1990, with ED50 about 1.5 mg/L. Circular dichroism and fluorescence spectrum analysis showed that the refolded sTRAIL had a structure similar to that of native protein with β-sheet secondary structure. This efficient procedure of sTRAIL renaturation may be useful for the mass production of this therapeutically important protein.

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42

Dolgikh, V. V., I. V. Senderskiy, G. V. Tetz e V. V. Tetz. "Optimization of the Protocol for the Isolation and Refolding of the Extracellular Domain of HER2 Expressed in Escherichia coli". Acta Naturae 6, n. 2 (15 giugno 2014): 106–9. http://dx.doi.org/10.32607/20758251-2014-6-2-106-109.

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Receptor 2 of the human epidermal growth factor (HER2/neu, c-erbB2) is a 185 kDa proto-oncogene protein characterized by an overexpression in some oncological diseases, including 30% of mammary glands cancers, as well as tumors in the ovary, stomach and other organs of the human body. Since HER2- tumor status testing is the essential part of a successful cancer treatment, the expression and purification of substantial amounts of the extracellular domain (ECD) of HER2 is an important task. The production of ECD HER2 in Escherichia coli has several advantages over the use of eukaryotic expression systems, but the bulk of the recombinant product in bacteria accumulates as insoluble protein inclusion bodies. In this study, we obtained ECD HER2 in Escherichia coli as insoluble inclusion bodies and elaborated a simple, efficient, and fast protocol for the solubilization, refolding, and isolation of the protein in soluble form.

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43

Awofisayo-Okuyelu, Adedoyin, Julii Brainard, Ian Hall e Noel McCarthy. "Incubation Period of Shiga Toxin–Producing Escherichia coli". Epidemiologic Reviews 41, n. 1 (2019): 121–29. http://dx.doi.org/10.1093/epirev/mxz001.

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Abstract Shiga toxin–producing Escherichia coli are pathogenic bacteria found in the gastrointestinal tract of humans. Severe infections could lead to life-threatening complications, especially in young children and the elderly. Understanding the distribution of the incubation period, which is currently inconsistent and ambiguous, can help in controlling the burden of disease. We conducted a systematic review of outbreak investigation reports, extracted individual incubation data and summary estimates, tested for heterogeneity, classified studies into subgroups with limited heterogeneity, and undertook a meta-analysis to identify factors that may contribute to the distribution of the pathogen’s incubation period. Twenty-eight studies were identified for inclusion in the review (1 of which included information on 2 outbreaks), and the resulting I2 value was 77%, indicating high heterogeneity. Studies were classified into 5 subgroups, with the mean incubation period ranging from 3.5 to 8.1 days. The length of the incubation period increased with patient age and decreased by 7.2 hours with every 10% increase in attack rate.

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44

Alimuddin, Alimuddin, Indra Lesmana, Agus Oman Sudrajat, Odang Carman e Irvan Faizal. "PRODUCTION AND BIOACTIVITY POTENTIAL OF THREE RECOMBINANT GROWTH HORMONES OF FARMED FISH". Indonesian Aquaculture Journal 5, n. 1 (30 giugno 2010): 11. http://dx.doi.org/10.15578/iaj.5.1.2010.11-17.

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This study was aimed to produce recombinant growth hormone (rGH) from giant grouper (Epinephelus lanceolatus), giant gouramy (Osphronemus gouramy) and common carp (Cyprinus carpio) and compare their bioactivity potential by means of inducing the growth hormone of juvenile Nile tilapia (Oreochromis niloticus) as the model. DNA fragment encoding mature GH protein of giant grouper (El-mGH), giant gouramy (Og-mGH) and common carp (Cc-mGH) was amplified by PCR method. The purified PCR products were ligated to pCold-1 to generate pCold/El-mGH, pCold/OgmGH, and pCold/Cc-mGH protein expression vector, respectively. Each of the expression vectors was transformed into the Escherichia coli BL21. E. coli BL21 was cultured using 2xYT medium and protein production was induced by cold shock at 15±1oC for overnight. The inclusion bodies of E. coli transformants containing protein expression vector were isolated by sonication method, and rGH production was analyzed by SDS-PAGE. Juvenile of Nile tilapia of average body weight of 12.41±3.28 g was intramuscularly injected once a week for 4 weeks with 1 μg inclusion body containing rGH per gram fish body weight. The result showed that rGH in molecular weight of about 25 kDa was obtained. Fish injected with rGH of El-mGH, Cc-mGH and Og-mGH grew 20.94%, 18.09%, and 16.99% faster, respectively, compared with the control. This result indicated that the three rGH produced in E. coli possessed biological activity when tested on Nile tilapia and further research is needed to find its effect on the growth of other aquaculture fish species.

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45

Fan, Gaofu, Zhiguo Yu, Jie Tang, Ruomeng Dai e Zhenguo Xu. "Preparation of gallic acid-hydroxypropyl-β-cyclodextrin inclusion compound and study on its effect mechanism on Escherichia coli in vitro". Materials Express 11, n. 5 (1 maggio 2021): 655–62. http://dx.doi.org/10.1166/mex.2021.1968.

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Abstract (sommario):

The purpose of this paper is to optimize the preparation process of gallic acid-hydroxypropyl-β-cyclodextrin inclusion compound and to study its antibacterial effect in vitro. Orthogonal test was used to screen the preparation method of the compound. The microscopic appearance of the compound was observed by microscope, and the solubility of the compound was detected by the dissolution method. The antibacterial activity of the compound was measured by the Oxford cup method. Under the transmission electron microscope, the microstructure of the cells and the intracellular ultrastructure changes were observed. The results showed that when the molar ratio of gallic acid and hydroxypropyl-β-cyclodextrin inclusion was 1:1, the ethanol concentration was 80%, the inclusion temperature was 30 °C, and the inclusion time was 0.5 h, the inclusion effect was the best with an inclusion rate of 99.45%; the solubility of gallic acid before the inclusion was (6.515 ±0.55) mg/mL, while after the inclusion, the solubility increased by more than 100 times to (710.048 ±1.08) mg/mL; gallic acid-hydroxypropyl-β-cyclodextrin inclusion compound could dissolve and destroy the cell membrane of Escherichia coli. The preparation process of gallic acid-hydroxypropyl-β-cyclodextrin inclusion compound was stable and feasible, and there was a good inclusion effect. After the inclusion, the solubility of gallic acid increased significantly; the prepared inclusion compound could inhibit the gram-negative bacteria Escherichia coli by destroying the integrity of the bacterial cell membrane, which could provide data support for the application and development of gallic acid.

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46

McCusker, Emily, e Anne Skaja Robinson. "Refolding of G protein α subunits from inclusion bodies expressed in Escherichia coli". Protein Expression and Purification 58, n. 2 (aprile 2008): 342–55. http://dx.doi.org/10.1016/j.pep.2007.11.015.

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47

Huang, Liurong, Haile Ma, Yunliang Li e Shuxiang Li. "Antihypertensive activity of recombinant peptide IYPR expressed in Escherichia coli as inclusion bodies". Protein Expression and Purification 83, n. 1 (maggio 2012): 15–20. http://dx.doi.org/10.1016/j.pep.2012.02.004.

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48

Valax, Pascal, e George Georgiou. "Molecular characterization of .beta.-lactamase inclusion bodies produced in Escherichia coli. 1. Composition". Biotechnology Progress 9, n. 5 (settembre 1993): 539–47. http://dx.doi.org/10.1021/bp00023a014.

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49

Georgiou, G., J. N. Telford, M. L. Shuler e D. B. Wilson. "Localization of inclusion bodies in Escherichia coli overproducing beta-lactamase or alkaline phosphatase." Applied and Environmental Microbiology 52, n. 5 (1986): 1157–61. http://dx.doi.org/10.1128/aem.52.5.1157-1161.1986.

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50

Sinacola, Jessica R., e Anne S. Robinson. "Rapid refolding and polishing of single-chain antibodies from Escherichia coli inclusion bodies". Protein Expression and Purification 26, n. 2 (novembre 2002): 301–8. http://dx.doi.org/10.1016/s1046-5928(02)00538-7.

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