Search results for: recombinant
#32957048 2020/09/15 To Up
Alleviating product inhibition of Trichoderma reesei cellulase complex with a product-activated mushroom endoglucanase.P
Gen Zou, Dapeng Bao, Ying Wang, Sichi Zhou, Meili Xiao, Zhanshan Yang, Yinmei Wang, Zhihua Zhou
2065 related Products with: Alleviating product inhibition of Trichoderma reesei cellulase complex with a product-activated mushroom endoglucanase.1 ml100 ul Product tipe: Anti0.2 mg50g25 µgcase50 ug Product tipe: Antib500g 100 TESTS250 ml1 L.50 ug Product tipe: Antib
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#32956955 2020/09/09 To Up
Production of a lyophilized ready-to-use yeast killer toxin with possible applications in the wine and food industries.Kpkt is a yeast killer toxin, naturally produced by Tetrapisispora phaffii, with possible applications in winemaking due to its antimicrobial activity on wine-related yeasts including Kloeckera/Hanseniaspora, Saccharomycodes and Zygosaccharomyces. Here, Kpkt coding gene was expressed in Komagataella phaffii (formerly Pichia pastoris) and the bioreactor production of the recombinant toxin (rKpkt) was obtained. Moreover, to produce a ready-to-use preparation of rKpkt, the cell-free supernatant of the K. phaffii recombinant killer clone was 80-fold concentrated and lyophilized. The resulting preparation could be easily solubilized in sterile distilled water and maintained its killer activity for up to six months at 4 °C. When applied to grape must, it exerted an extensive killer activity on wild wine-related yeasts while proving compatible with the fermentative activity of actively growing Saccharomyces cerevisiae starter strains. Moreover, it displayed a strong microbicidal effect on a variety of bacterial species including lactic acid bacteria and food-borne pathogens. On the contrary it showed no lethal effect on filamentous fungi and on Ceratitis capitata and Musca domestica, two insect species that may serve as non-mammalian model for biomedical research. Based on these results, bioreactor production and lyophilization represent an interesting option for the exploitation of this killer toxin that, due to its spectrum of action, may find application in the control of microbial contaminations in the wine and food industries.
Gavino Carboni, Francesco Fancello, Giacomo Zara, Severino Zara, Luca Ruiu, Ivana Marova, Giovanni Pinna, Marilena Budroni, Ilaria Mannazzu
2858 related Products with: Production of a lyophilized ready-to-use yeast killer toxin with possible applications in the wine and food industries.1100ug Lyophilized100ug Lyophilized100ug Lyophilized100ug Lyophilized100ug Lyophilized100ug Lyophilized100ug Lyophilized100ug Lyophilized100ug Lyophilized100ug Lyophilized
#32956801 2020/09/18 To Up
Characterization of the Fc-III-4C-based Recombinant Protein Expression System by Using Carbonic Anhydrase as the Model Protein.Development of new affinity tags is important for recombinant protein expression and purification. Based on our earlier work, we devised an affinity tag by addition of two cysteine residues onto the N- and C-termini of the Fc-III peptide and designated as the Fc-III-4C tag, in which four cysteine residues form two disulfide linkages. The binding affinity of Fc-III-4C tag to human IgG is measured as 2.28 nM (K) and is 100 times higher than that of the Fc-III tag to IgG. Fc-III-4C tagged carbonic anhydrase (CA) can be effectively purified with IgG-immobilized beads, and Fc-III-4C tag does not possess adverse effects on the structure and stability of CA. Furthermore, the Fc-III-4C tagged protein binds to multiple transition metal ions, which enhances activities of enzymes that use metal ions as co-factors. These results suggest that Fc-III-4C tag is a useful tool for expression and purification of recombinant proteins and enhances the activities of some fusion proteins that use Zn or Cu as cofactors.
Yiyi Gong, Meiqi Yi, Lin Zhang, Shan Feng, Haiteng Deng
2209 related Products with: Characterization of the Fc-III-4C-based Recombinant Protein Expression System by Using Carbonic Anhydrase as the Model Protein.2x 100ug100 U21mg10100 ug25 ul1mg25 ug1mg50 Test5 ug
#32956771 2020/09/18 To Up
Identification and characterization of a cDNA encoding a gametocyte-specific protein of the avian coccidial parasite Eimeria necatrix.Gametocyte proteins of Eimeria spp. are essential components of the oocyst wall, and some of these proteins have been analysed to identify targets of transmission-blocking vaccines against avian coccidiosis. In the present study, a cDNA from E. necatrix gametocytes was cloned and sequenced. The cDNA is 1,473 bp in length and encodes a 490-amino-acid protein containing a tyrosine-serine (Tyr/Ser)-rich domain and a proline-methionine (Pro/Met)-rich domain. A quantitative real-time PCR (qPCR) analysis showed that the cDNA is expressed only during gametogenesis. A fragment containing the Tyr/Ser-rich domain (rEnGAM59) was expressed in Escherichia coli BL21 (DE3) cells. Immunoblotting showed that rEnGAM59 was recognized by the serum of convalescent chickens after infection with E. necatrix, and that an anti-rEnGAM59 antibody recognized a ∼59 kDa protein and two other proteins (∼35 kDa and ∼33 kDa) in gametocyte extracts. An immunofluorescence assay showed that the anti-rEnGAM59 antibody recognized wall-forming bodies in the macrogametocytes and oocyst walls. An in vivo vaccination and challenge trial was conducted to test the potential utility of rEnGAM59 as a vaccine. Immunized chickens performed better than the unimmunized and challenged (positive control) chickens. The intestinal lesion scores were significantly lower in the immunized groups than in the positive control group (P < 0.05). In contrast, the body weight gains (BWG) were significantly higher in the immunized groups than in the positive control group (P < 0.05). There were no significant differences in the lesion scores and BWG between the groups immunized with rEnGAM59 protein or with live oocysts (P > 0.05). Chickens immunized with rEnGAM59 protein had a significantly higher antigen-specific serum IgY response (P < 0.05). rEnGAM59 protein can be used as candidate antigen to develop a recombinant coccidiosis vaccine.
Dandan Liu, Feiyan Wang, Liqin Cao, Lele Wang, Shijie Su, Zhaofeng Hou, Jinjun Xu, Junjie Hu, Jianping Tao
1361 related Products with: Identification and characterization of a cDNA encoding a gametocyte-specific protein of the avian coccidial parasite Eimeria necatrix.100ug1000 TESTS/0.65ml100ul0.05 mg2 Pieces/Box1 mg50 100.00 ug10ìg0.1 mg
#32955862 2020/09/21 To Up
Tuning the Structural Integrity and Mechanical Properties of Globular Protein Vesicles by Blending Crosslinkable and Non-Crosslinkable Building Blocks.Vesicles made from functionally folded, globular proteins that perform specific biological activities, such as catalysis, sensing, or therapeutics, allow for potential applications as artificial cells, micro-bioreactors, or protein drug delivery. The mechanical properties of vesicle membranes, including elastic modulus and hardness, play a critical role in dictating the stability and shape transformation of the vesicles under external stimuli-triggers. Herein, we have developed a strategy to tune the mechanical properties and integrity of globular protein vesicle (GPV) membranes of which building molecules are recombinant fusion protein complexes: mCherry fused with acidic leucine zipper (mCherry-Z) and basic leucine zipper fused with elastin-like polypeptide (Z-ELP). To control the mechanical properties of GPVs, we introduced a nonstandard amino acid (para-azidophenylalanine (pAzF)) into the ELP domains (ELP-X), which enabled the creation of crosslinked vesicles under ultraviolet (UV) irradiation. Crosslinked GPVs made from mCherry-Z/Z-ELP-X complexes presented higher stability than non-crosslinked GPVs under hypotonic osmotic stress. The degree of swelling of GPVs increased as less crosslinking was achieved in the vesicle membranes, which resulted in disassembly of GPVs into membraneless coacervates. Nanoindentation by atomic force microscopy (AFM) confirmed that stiffness and Young's elastic modulus of GPVs increase as the blending molar ratio of Z-ELP-X to Z-ELP increases to make vesicles. The results obtained in this study suggest a rational design to make GPVs with tunable mechanical properties for target applications by simply varying the blending ratio of Z-ELP and Z-ELP-X in the vesicle self-assembly.
Ruwen Tan, Jooyong Shin, Jiwoong Heo, Blair Cole, Jinkee Hong, Yeongseon Jang
1340 related Products with: Tuning the Structural Integrity and Mechanical Properties of Globular Protein Vesicles by Blending Crosslinkable and Non-Crosslinkable Building Blocks.1000 TESTS/0.65ml100ul100ug500 mg100ug25 mg210 mg25 mg 5 G 5 G1000
#32955827 2020/09/21 To Up
Tagraxofusp and anti-CD123 in blastic plasmacytoid dendritic cell neoplasm: a new hope.Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare hematological malignancy, characterized by poor prognosis if treated with conventional therapy. Allogenic hematologic stem cell transplant can improve survival and can be curative but it is available in a small percentage of patients given that the median age at diagnosis is 70 years. In this scenario it is assumed that only the development of precision medicine-driven therapy will change BPDCN patient prognosis. CD123 (the α-subunit of interleukin (IL)-3 receptor) is over-expressed on BPDCN cells surface and seems to be the ideal marker to develop antibody-based therapies. Tagraxofusp (Elzonris), a recombinant immunotoxin consisting of human interleukin-3 fused to a truncated diphtheria toxin, has been approved by FDA in December 2018 for the treatment of BPDCN in adult and pediatric patients. Tagraxofusp has shown promising clinical activity, with a high overall response rate and quite manageable safety profile even in elderly patients. It seems to improve overall survival too but comparative trials are necessary to confirm this. Adverse events are commonly reported and the most important are transaminitis, thrombocytopenia and capillary leak syndrome (CLS). Therefore to prevent the onset of severe CLS is recommended to reserve tagraxofusp for patients with preserved hepatic and cardiac functions, and to strictly observe serum albumin level. Further studies are required to resolve many several unanswered questions about tagraxofusp. In this review, we will resume and discuss pharmacological characteristic of tagraxofusp, results of clinical trials leading to its approval by FDA in 2018 and future perspectives about its use in BPDCN and other hematological malignancies.
Delia Cangini, Paolo Silimbani, Alessandro Cafaro, Maria Benedetta Giannini, Carla Masini, Andrea Ghelli Luserna Di Rorá, Giorgia Simonetti, Giovanni Martinelli, Claudio Cerchione
2998 related Products with: Tagraxofusp and anti-CD123 in blastic plasmacytoid dendritic cell neoplasm: a new hope.2 ml100 μg100 μg100ug Lyophilized100 µg100ug Lyophilized100ug Lyophilized200 100ug Lyophilized1 mL100ug Lyophilized
#32955754 2020/09/21 To Up
InteBac - An integrated bacterial and baculovirus expression vector suite.The successful production of recombinant protein for biochemical, biophysical and structural biological studies critically depends on the correct expression organism. Currently the most commonly used expression organisms for structural studies are E. coli (ca. 70% of all PDB structures) and the baculovirus/ insect cell expression system (ca. 5% of all PDB structures). While insect cell expression is frequently successful for large eukaryotic proteins, it is relatively expensive and time consuming compared to E. coli expression. Frequently the decision to carry out a baculovirus project means restarting cloning from scratch. Here we describe an integrated system that allows the simultaneous cloning into E. coli and baculovirus expression vectors using the same PCR products. The system offers a flexible array of N- and C- terminal affinity, solublisation and utility tags, and the speed allows expression screening to be completed in E. coli, before carrying out time and cost intensive experiments in baculovirus. Importantly, we describe a means of rapidly generating polycistronic bacterial constructs based on the hugely successful biGBac system, making InteBac of particular interest for researchers working on recombinant protein complexes. This article is protected by copyright. All rights reserved.
Veronika Altmannova, Andreas Blaha, Susanne Astrinidis, Heidi Reichle, John R Weir
2563 related Products with: InteBac - An integrated bacterial and baculovirus expression vector suite.1 mg100ul
#32955743 2020/09/21 To Up
Impact of peri-procedural subcutaneous parathyroid hormone on control of hypocalcemia in APS-1/APECED patients undergoing invasive procedures.The monogenic disorder autoimmune polyendocrine syndrome type 1 (APS-1) or autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) manifests frequently with hypoparathyroidism, which requires treatment with oral supplementation with calcium and active vitamin D analogs. The majority of APS-1/APECED patients also suffer from intestinal malabsorption, which complicates the management of hypoparathyroidism and may lead to refractory severe hypocalcemia. In such situations, reliance on intravenous calcium carries a high risk of nephrocalcinosis and renal damage.
Karen K Winer, Monica M Schmitt, Elise M N Ferre, Kevin P Fennelly, Kenneth N Olivier, Theo Heller, Michail S Lionakis
1674 related Products with: Impact of peri-procedural subcutaneous parathyroid hormone on control of hypocalcemia in APS-1/APECED patients undergoing invasive procedures.96 tests100ug
#32955164 2020/09/21 To Up
Hematological adaptations and detection of recombinant human erythropoietin combined with chronic hypoxia.This study evaluated whether recombinant human erythropoietin (rhEpo) treatment combined with chronic hypoxia provided an additive erythropoietic response and whether the Athlete Biological Passport (ABP) sensitivity improved with hypoxia. Two interventions were completed, each containing four weeks baseline, four weeks exposure at sea-level or 2,320m of altitude and four weeks follow-up. Participants were randomly assigned to 20 IU·kg bw rhEpo or placebo injections every second day for three weeks during the exposure period at sea-level (rhEpo n=25, placebo n=9) or at altitude (rhEpo n=12, placebo n=27). Venous blood was analyzed weekly. Combining rhEpo and hypoxia induced larger changes compared with rhEpo or hypoxia alone for [Hb] (P<0.001, P>0.05, respectively), reticulocyte percentage (P<0.001) and OFF-hr score (P<0.01, P<0.001, respectively). The most pronounced effect was observed for reticulocyte percentage with up to ~35% (P<0.001) and ~45% (P<0.001) higher levels compared with rhEpo or hypoxia only, respectively. The ABP sensitivity for the combined treatment was 54 and 35 percentage points higher for [Hb] (P<0.05) and reticulocyte percentage (P<0.05), respectively, but similar for OFF-hr score, compared with rhEpo at sea-level. Across any time point, [Hb] and OFF-hr score combined identified 14 unique true-positive participants (56%) at sea-level and 12 unique true-positive participants (100%) at altitude. However, a concurrent reduction in specificity existed at altitude. In conclusion, rhEpo treatment combined with hypoxic exposure provided an additive erythropoietic response compared to rhEpo or hypoxic exposure alone. Correspondingly, ABP was more sensitive to rhEpo at altitude than at sea-level, but a compromised specificity existed with hypoxic exposure.
Jacob Bejder, Andreas Breenfeldt Andersen, Thomas Christian Bonne, Jesper Linkis, Niels Vidiendal Olsen, Jesús Rodríguez Huertas, Nikolai Baastrup Nordsborg
2071 related Products with: Hematological adaptations and detection of recombinant human erythropoietin combined with chronic hypoxia.100ul1 mg1 mg1mg100μg/vial510020 μg2551 mg100
#32955147 2020/09/21 To Up
Rapid sequence modification in the highly polymorphic region (HPR) of the hemagglutinin gene of the infectious salmon anaemia virus (ISAV) suggests intra-segmental template switching recombination.The ISAV has a genome composed of eight segments of (-)ssRNA, segment 6 codes for the hemagglutinin-esterase protein, and has the most variable region of the genome, the highly polymorphic region (HPR), which is unique among orthomyxoviruses. The HPR has been associated with virulence, infectivity and pathogenicity. The full length of the HPR is called HPR0 and the strain with this HPR is avirulent, in contrast to strains with deleted HPR that are virulent to varying degrees. The molecular mechanism that gives rise to the different HPRs remains unclear. Here, we studied in vitro the evolution of reassortant recombinant ISAV (rISAV) in Atlantic salmon head kidney (ASK) cells. To this end, we rescued and cultivated a set of rISAV with different segment 6-HPR genotypes using a reverse genetics system and then sequencing HPR regions of the viruses. Our results show rapid multiple recombination events in ISAV, with sequence insertions and deletions in the HPR, indicating a dynamic process. Inserted sequences can be found in four segments of the ISAV genome (segments 1, 5, 6, and 8). The results suggest intra-segmental heterologous recombination, probably by class I and class II template switching, similar to the proposed segment 5 recombination mechanism.
Matías Cárdenas, Claudia Galleguillos, Karina Acevedo, Catarina Ananias, Javiera Alarcón, Sofía Michelson, Jorge Toledo, Margarita Montoya, Claudio Meneses, Eduardo Castro-Nallar, Yesseny Vásquez-Martínez, Marcelo Cortez-San Martin
1597 related Products with: Rapid sequence modification in the highly polymorphic region (HPR) of the hemagglutinin gene of the infectious salmon anaemia virus (ISAV) suggests intra-segmental template switching recombination.11min 2 cartons100.00 ul
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