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BMC Biotechnology

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https://www.readbyqxmd.com/read/28100213/transcriptomic-response-of-wolf-spider-pardosa-pseudoannulata-to-transgenic-rice-expressing-bacillus-thuringiensis-cry1ab-protein
#1
Juan Wang, Yuande Peng, Kaifu Xiao, Baoyang Wei, Jilin Hu, Zhi Wang, Qisheng Song, Xuguo Zhou
BACKGROUND: Bacillum thuringiensis (Bt) toxin produced in Cry1-expressing genetically modified rice (Bt rice) is highly effective to control lepidopteran pests, which reduces the needs for synthetic insecticides. Non-target organisms can be exposed to Bt toxins through direct feeding or trophic interactions in the field. The wolf spider Pardosa pseudoannulata, one of the dominant predators in South China, plays a crucial role in the rice agroecosystem. In this study, we investigated transcriptome responses of the 5th instar spiders fed on preys maintained on Bt- and non-Bt rice...
January 18, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28100205/a-fast-and-effective-determination-of-the-biodistribution-and-subcellular-localization-of-fluorescent-immunoliposomes-in-freshly-excised-animal-organs
#2
Felista L Tansi, Ronny Rüger, Ansgar M Kollmeier, Claudia Böhm, Roland E Kontermann, Ulf K Teichgraeber, Alfred Fahr, Ingrid Hilger
BACKGROUND: Preclinical research implementing fluorescence-based approaches is inevitable for drug discovery and technology. For example, a variety of contrast agents developed for biomedical imaging are usually evaluated in cell systems and animal models based on their conjugation to fluorescent dyes. Biodistribution studies of excised organs are often performed by macroscopic imaging, whereas the subcellular localization though vital, is often neglected or further validated by histological procedures...
January 18, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28095828/improved-genome-sequencing-using-an-engineered-transposase
#3
Amirali Kia, Christian Gloeckner, Trina Osothprarop, Niall Gormley, Erin Bomati, Michelle Stephenson, Igor Goryshin, Molly Min He
BACKGROUND: Next-generation sequencing (NGS) has transformed genomic research by reducing turnaround time and cost. However, no major breakthrough has been made in the upstream library preparation methods until the transposase-based Nextera method was invented. Nextera combines DNA fragmentation and barcoding in a single tube reaction and therefore enables a very fast workflow to sequencing-ready DNA libraries within a couple of hours. When compared to the traditional ligation-based methods, transposed-based Nextera has a slight insertion bias...
January 17, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28088197/production-of-human-pro-relaxin-h2-in-the-yeast-pichia-pastoris
#4
D Cimini, K Della Corte, R Finamore, L Andreozzi, A Stellavato, A V A Pirozzi, F Ferrara, R Formisano, M De Rosa, M Chino, L Lista, A Lombardi, V Pavone, C Schiraldi
BACKGROUND: Initially known as the reproductive hormone, relaxin was shown to possess other therapeutically useful properties that include extracellular matrix remodeling, anti-inflammatory, anti-ischemic and angiogenic effects. All these findings make relaxin a potential drug for diverse medical applications. Its precursor, pro-relaxin, is an 18 kDa protein, that shows activity in in vitro assays. Since extraction of relaxin from animal tissues raises several issues, prokaryotes and eukaryotes were both used as expression systems for recombinant relaxin production...
January 14, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28088196/the-antibacterial-activity-and-mechanism-of-ginkgolic-acid-c15-1
#5
Zhebin Hua, Caie Wu, Gongjian Fan, Zhenxing Tang, Fuliang Cao
BACKGROUND: The present study investigated the antibacterial activity and underlying mechanisms of ginkgolic acid (GA) C15:1 monomer using green fluorescent protein (GFP)-labeled bacteria strains. RESULTS: GA presented significant antibacterial activity against Gram-positive bacteria but generally did not affect the growth of Gram-negative bacteria. The studies of the antibacterial mechanism indicated that large amounts of GA (C15:1) could penetrate GFP-labeled Bacillus amyloliquefaciens in a short period of time, and as a result, led to the quenching of GFP in bacteria...
January 14, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28081707/antigen-specific-single-b-cell-sorting-and-expression-cloning-from-immunoglobulin-humanized-rats-a-rapid-and-versatile-method-for-the-generation-of-high-affinity-and-discriminative-human-monoclonal-antibodies
#6
Laure-Hélène Ouisse, Laetitia Gautreau-Rolland, Marie-Claire Devilder, Michael Osborn, Melinda Moyon, Jonathan Visentin, Frank Halary, Marianne Bruggemann, Roland Buelow, Ignacio Anegon, Xavier Saulquin
BACKGROUND: There is an ever-increasing need of monoclonal antibodies (mAbs) for biomedical applications and fully human binders are particularly desirable due to their reduced immunogenicity in patients. We have applied a strategy for the isolation of antigen-specific B cells using tetramerized proteins and single-cell sorting followed by reconstruction of human mAbs by RT-PCR and expression cloning. RESULTS: This strategy, using human peripheral blood B cells, enabled the production of low affinity human mAbs against major histocompatibility complex molecules loaded with peptides (pMHC)...
January 9, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28061848/multi-antigen-avian-influenza-a-h7n9-virus-like-particles-particulate-characterizations-and-immunogenicity-evaluation-in-murine-and-avian-models
#7
Che-Ming Jack Hu, Chu-Yang Chien, Ming-Tsan Liu, Zih-Syun Fang, Sui-Yuan Chang, Rong-Huay Juang, Shih-Chung Chang, Hui-Wen Chen
BACKGROUND: Human infection with avian influenza A virus (H7N9) was first reported in China in March 2013. Since then, hundreds of cases have been confirmed showing severe symptoms with a high mortality rate. The virus was transmitted from avian species to humans and has spread to many neighboring areas, raising serious concerns over its pandemic potential. Towards containing the disease, the goal of this study is to prepare a virus-like particle (VLP) that consists of hemagglutinin (HA), neuraminidase (NA) and matrix protein 1 (M1) derived from the human isolate A/Taiwan/S02076/2013(H7N9) for potential vaccine development...
January 7, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28056928/new-ligation-independent-cloning-vectors-for-expression-of-recombinant-proteins-with-a-self-cleaving-cpd-6xhis-tag
#8
Marco Biancucci, Jazel S Dolores, Jennifer Wong, Sarah Grimshaw, Wayne F Anderson, Karla J F Satchell, Keehwan Kwon
BACKGROUND: Recombinant protein purification is a crucial step for biochemistry and structural biology fields. Rapid robust purification methods utilize various peptide or protein tags fused to the target protein for affinity purification using corresponding matrices and to enhance solubility. However, affinity/solubility-tags often need to be removed in order to conduct functional and structural studies, adding complexities to purification protocols. RESULTS: In this work, the Vibrio cholerae MARTX toxin Cysteine Protease Domain (CPD) was inserted in a ligation-independent cloning (LIC) vector to create a C-terminal 6xHis-tagged inducible autoprocessing enzyme tag, called "the CPD-tag"...
January 5, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/27955656/means-of-enhancing-bone-fracture-healing-optimal-cell-source-isolation-methods-and-acoustic-stimulation
#9
Corina Adriana Ghebes, Maaike Vera Jasmijn Braham, Adelgunde Veronica Clemens Maria Zeegers, Auke Jan Sijbe Renard, Hugo Fernandes, Daniel B F Saris
BACKGROUND: The human body has an extensive capacity to regenerate bone tissue after trauma. However large defects such as long bone fractures of the lower limbs cannot be restored without intervention and often lead to nonunion. Therefore, the aim of the present study was to assess the pool and biological functions of human mesenchymal stromal cells (hMSCs) isolated from different bone marrow locations of the lower limbs and to identify novel strategies to prime the cells prior to their use in bone fracture healing...
December 12, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27938351/modular-assembly-of-synthetic-proteins-that-span-the-plasma-membrane-in-mammalian-cells
#10
Anam Qudrat, Kevin Truong
BACKGROUND: To achieve synthetic control over how a cell responds to other cells or the extracellular environment, it is important to reliably engineer proteins that can traffic and span the plasma membrane. Using a modular approach to assemble proteins, we identified the minimum necessary components required to engineer such membrane-spanning proteins with predictable orientation in mammalian cells. RESULTS: While a transmembrane domain (TM) fused to the N-terminus of a protein is sufficient to traffic it to the endoplasmic reticulum (ER), an additional signal peptidase cleavage site downstream of this TM enhanced sorting out of the ER...
December 9, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27908283/expression-of-a-fungal-manganese-peroxidase-in-escherichia-coli-a-comparison-between-the-soluble-and-refolded-enzymes
#11
Nan Wang, Kai Ren, Rong Jia, Wenting Chen, Ruirui Sun
BACKGROUND: Manganese peroxidase (MnP) from Irpex lacteus F17 has been shown to have a strong ability to degrade recalcitrant aromatic pollutants. In this study, a recombinant MnP from I. lacteus F17 was expressed in Escherichia coli Rosetta (DE3) in the form of inclusion bodies, which were refolded to achieve an active enzyme. Further, we optimized the in vitro refolding conditions to increase the recovery yield of the recombinant protein production. Additionally, we attempted to express recombinant MnP in soluble form in E...
December 1, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27905914/development-of-a-gene-synthesis-platform-for-the-efficient-large-scale-production-of-small-genes-encoding-animal-toxins
#12
Ana Filipa Sequeira, Joana L A Brás, Catarina I P D Guerreiro, Renaud Vincentelli, Carlos M G A Fontes
BACKGROUND: Gene synthesis is becoming an important tool in many fields of recombinant DNA technology, including recombinant protein production. De novo gene synthesis is quickly replacing the classical cloning and mutagenesis procedures and allows generating nucleic acids for which no template is available. In addition, when coupled with efficient gene design algorithms that optimize codon usage, it leads to high levels of recombinant protein expression. RESULTS: Here, we describe the development of an optimized gene synthesis platform that was applied to the large scale production of small genes encoding venom peptides...
December 1, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27903244/cryopreservation-of-dermal-fibroblasts-and-keratinocytes-in-hydroxyethyl-starch-based-cryoprotectants
#13
Yahaira Naaldijk, Adiv A Johnson, Annett Friedrich-Stöckigt, Alexandra Stolzing
BACKGROUND: Preservation of human skin fibroblasts and keratinocytes is essential for the creation of skin tissue banks. For successful cryopreservation of cells, selection of an appropriate cryoprotectant agent (CPA) is imperative. The aim of this study was to identify CPAs that minimize toxic effects and allow for the preservation of human fibroblasts and keratinocytes in suspension and in monolayers. RESULTS: We cryopreserved human fibroblasts and keratinocytes with different CPAs and compared them to fresh, unfrozen cells...
December 1, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27881117/novel-prokaryotic-expression-of-thioredoxin-fused-insulinoma-associated-protein-tyrosine-phosphatase-2-ia-2-its-characterization-and-immunodiagnostic-application
#14
Luciano Lucas Guerra, Natalia Inés Faccinetti, Aldana Trabucchi, Bruno David Rovitto, Adriana Victoria Sabljic, Edgardo Poskus, Ruben Francisco Iacono, Silvina Noemí Valdez
BACKGROUND: The insulinoma associated protein tyrosine phosphatase 2 (IA-2) is one of the immunodominant autoantigens involved in the autoimmune attack to the beta-cell in Type 1 Diabetes Mellitus. In this work we have developed a complete and original process for the production and recovery of the properly folded intracellular domain of IA-2 fused to thioredoxin (TrxIA-2ic) in Escherichia coli GI698 and GI724 strains. We have also carried out the biochemical and immunochemical characterization of TrxIA-2icand design variants of non-radiometric immunoassays for the efficient detection of IA-2 autoantibodies (IA-2A)...
November 24, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27876044/generation-of-monoclonal-antibodies-against-native-viral-proteins-using-antigen-expressing-mammalian-cells-for-mouse-immunization
#15
Natalie Rose, Carlos Augusto Pinho-Nascimento, Alessia Ruggieri, Paola Favuzza, Marco Tamborrini, Hanna Roth, Marcia Terezinha Baroni de Moraes, Hugues Matile, Thomas Jänisch, Gerd Pluschke, Katharina Röltgen
BACKGROUND: Due to their rising incidence and progressive geographical spread, infections with mosquito-borne viruses, such as dengue (DENV), chikungunya and zika virus, have developed into major public health challenges. Since all of these viruses may cause similar symptoms and can occur in concurrent epidemics, tools for their differential diagnosis and epidemiological monitoring are of urgent need. RESULTS: Here we report the application of a novel strategy to rapidly generate monoclonal antibodies (mAbs) against native viral antigens, exemplified for the DENV nonstructural glycoprotein 1 (NS1)...
November 22, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27871321/biochemical-characterization-and-synergism-of-cellulolytic-enzyme-system-from-chaetomium-globosum-on-rice-straw-saccharification
#16
Wanwitoo Wanmolee, Warasirin Sornlake, Nakul Rattanaphan, Surisa Suwannarangsee, Navadol Laosiripojana, Verawat Champreda
BACKGROUND: Efficient hydrolysis of lignocellulosic materials to sugars for conversion to biofuels and chemicals is a key step in biorefinery. Designing an active saccharifying enzyme system with synergy among their components is considered a promising approach. RESULTS: In this study, a lignocellulose-degrading enzyme system of Chaetomium globosum BCC5776 (CG-Cel) was characterized for its activity and proteomic profiles, and synergism with accessory enzymes. The highest cellulase productivity of 0...
November 21, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27871279/biodegradation-of-lignin-and-nicotine-with-white-rot-fungi-for-the-delignification-and-detoxification-of-tobacco-stalk
#17
Yulong Su, He Xian, Sujuan Shi, Chengsheng Zhang, S M Nuruzzaman Manik, Jingjing Mao, Ge Zhang, Weihong Liao, Qian Wang, Haobao Liu
BACKGROUND: Tobacco stalk is one kind of abundant crop residues in China. The high lignification of tobacco stalk increases its reusing cost and the existing of nicotine will cause serious pollution. The biodegradation of lignocellulosic biomass has been demonstrated to be an environmental and economical approach for the utilization of plant stalk. Meanwhile, many nicotine-degrading microorganisms were found in nature. However, microorganisms which could degraded both nicotine and lignin haven't been reported...
November 21, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27855668/identification-and-characterization-of-two-new-5-keto-4-deoxy-d-glucarate-dehydratases-decarboxylases
#18
André Pick, Barbara Beer, Risa Hemmi, Rena Momma, Jochen Schmid, Kenji Miyamoto, Volker Sieber
BACKGROUND: Hexuronic acids such as D-galacturonic acid and D-glucuronic acid can be utilized via different pathways within the metabolism of microorganisms. One representative, the oxidative pathway, generates α-keto-glutarate as the direct link entering towards the citric acid cycle. The penultimate enzyme, keto-deoxy glucarate dehydratase/decarboxylase, catalyses the dehydration and decarboxylation of keto-deoxy glucarate to α-keto-glutarate semialdehyde. This enzymatic reaction can be tracked continuously by applying a pH-shift assay...
November 17, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27852252/a-novel-pyruvate-kinase-and-its-application-in-lactic-acid-production-under-oxygen-deprivation-in-corynebacterium-glutamicum
#19
Xin Chai, Xiuling Shang, Yu Zhang, Shuwen Liu, Yong Liang, Yun Zhang, Tingyi Wen
BACKGROUND: Pyruvate kinase (Pyk) catalyzes the generation of pyruvate and ATP in glycolysis and functions as a key switch in the regulation of carbon flux distribution. Both the substrates and products of Pyk are involved in the tricarboxylic acid cycle, anaplerosis and energy anabolism, which places Pyk at a primary metabolic intersection. Pyks are highly conserved in most bacteria and lower eukaryotes. Corynebacterium glutamicum is an industrial workhorse for the production of various amino acids and organic acids...
November 16, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27829414/repair-of-cartilage-defects-in-osteoarthritis-rats-with-induced-pluripotent-stem-cell-derived-chondrocytes
#20
Yanxia Zhu, Xiaomin Wu, Yuhong Liang, Hongsheng Gu, Kedong Song, Xuenong Zou, Guangqian Zhou
BACKGROUND: The incapacity of articular cartilage (AC) for self-repair after damage ultimately leads to the development of osteoarthritis. Stem cell-based therapy has been proposed for the treatment of osteoarthritis (OA) and induced pluripotent stem cells (iPSCs) are becoming a promising stem cell source. RESULTS: Three steps were developed to differentiate human iPSCs into chondrocytes which were transplanted into rat OA models induced by monosodium iodoacetate (MIA)...
November 9, 2016: BMC Biotechnology
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