1,335 results match your criteria BMC Biotechnology [Journal]


Tetanus toxin fragments and Bcl-2 fusion proteins: cytoprotection and retrograde axonal migration.

BMC Biotechnol 2018 Jun 11;18(1):39. Epub 2018 Jun 11.

Division of Neurology, Department of Brain and Neurosciences, Faculty of Medicine, Tottori University, Nishi-cho 36-1, Yonago, 683-8504, Japan.

Background: Tetanus neurotoxin (TeNT) is taken up at nerve terminals and undergoes retrograde migration. The toxic properties of TeNT reside in the toxin light chain (L), but like complete TeNT, the TeNT heavy chain (TTH) and the C-terminal domain (TTC) alone can bind and enter into neurons. Here, we explored whether atoxic fragments of TeNT could act as drug delivery vehicles in neurons. Read More

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June 2018
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DNA assembly with error correction on a droplet digital microfluidics platform.

BMC Biotechnol 2018 Jun 1;18(1):37. Epub 2018 Jun 1.

Stanford Genome Technology Center, Stanford University, 3165 Porter Drive, Palo Alto, CA, 94304, USA.

Background: Custom synthesized DNA is in high demand for synthetic biology applications. However, current technologies to produce these sequences using assembly from DNA oligonucleotides are costly and labor-intensive. The automation and reduced sample volumes afforded by microfluidic technologies could significantly decrease materials and labor costs associated with DNA synthesis. Read More

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Heterologous expression and biochemical characterization of a GHF9 endoglucanase from the termite Reticulitermes speratus in Pichia pastoris.

BMC Biotechnol 2018 Jun 1;18(1):35. Epub 2018 Jun 1.

National Key Laboratory of Biochemical Engineering, National Engineering Research Center for Biotechnology (Beijing), Key Laboratory of Biopharmaceutical Production & Formulation Engineering, PLA, Institute of Process Engineering, Chinese Academy of Sciences, No. 1 North 2nd Street, Beijing, 100190, China.

Background: Cellulases are of great significance for full utilization of lignocellulosic biomass. Termites have an efficient ability to degrade cellulose. Heterologous production of the termite-origin cellulases is the first step to realize their industrial applications. Read More

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Engineering Bacillus pumilus alkaline serine protease to increase its low-temperature proteolytic activity by directed evolution.

BMC Biotechnol 2018 Jun 1;18(1):34. Epub 2018 Jun 1.

Key Laboratory for Bio-resources and Eco-environment of Ministry of Education, Sichuan Key Laboratory of Molecular Biology and Biotechnology, College of Life Sciences, Sichuan University, Chengdu, Sichuan, 610064, People's Republic of China.

Background: Mesophilic alkaline serine proteases from various bacteria have been commercially applied in a range of industries owing to their high catalytic efficiency and wide substrate specificity. However, these proteases have an optimal catalytic temperature of approximately 50 °C, and their activity decreases significantly at low temperature. Therefore, to enhance their cold activity, it is necessary to improve the catalytic performance of these proteases at low temperature. Read More

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The importance of genotype identity, genetic heterogeneity, and bioinformatic handling for properly assessing genomic variation in transgenic plants.

BMC Biotechnol 2018 Jun 1;18(1):38. Epub 2018 Jun 1.

Bioinformatics and Computational Biology Program, University of Minnesota, Minneapolis, MN, USA.

Background: The advent of -omics technologies has enabled the resolution of fine molecular differences among individuals within a species. DNA sequence variations, such as single nucleotide polymorphisms or small deletions, can be tabulated for many kinds of genotype comparisons. However, experimental designs and analytical approaches are replete with ways to overestimate the level of variation present within a given sample. Read More

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Enhancing thermal tolerance of Aspergillus niger PhyA phytase directed by structural comparison and computational simulation.

BMC Biotechnol 2018 Jun 1;18(1):36. Epub 2018 Jun 1.

School of Life Sciences, Yunnan Normal University, Kunming, 650500, China.

Background: Phytase supplied in feeds for monogastric animals is important for improving nutrient uptake and reducing phosphorous pollution. High-thermostability phytases are particularly desirable due to their ability to withstand transient high temperatures during feed pelleting procedures. A comparison of crystal structures of the widely used industrial Aspergillus niger PhyA phytase (AnP) with its close homolog, the thermostable Aspergillus fumigatus phytase (AfP), suggests 18 residues in three segments associated with thermostability. Read More

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June 2018
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Preparation of an epitope-based recombinant diagnostic antigen specific to anti-phospholipase A receptor 1 antibodies.

BMC Biotechnol 2018 May 29;18(1):31. Epub 2018 May 29.

Jiangxi Province Key Laboratory for Laboratory Medicine, Department of Clinical Laboratory, The Second Affiliated Hospital of Nanchang University, Nanchang, 330006, China.

Background: According to recent studies, the phospholipase A2 receptor 1 (PLAR1) may be used as a biomarker to diagnose idiopathic membranous nephropathy (iMN). Moreover, the immune-dominant regions of PLAR1 have been identified. The aim of the present study was to construct a diagnostic antigen based on the immune-dominant region of PLAR1 and develop a specific serological detection method for PLAR1 antibodies. Read More

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FGF23 improves diabetic nephropathy by attenuating renal fibrosis and inflammation.

BMC Biotechnol 2018 May 30;18(1):33. Epub 2018 May 30.

Department of Biopharmacy, School of Pharmacy, Wenzhou Medical University, Chashan Town, Wenzhou, 325035, China.

Background: High level of serum fibroblast growth factor 23 (FGF23) is implicated in the development and progression of diabetic nephropathy (DN), making it a crucial factor in the pathogenesis of DN. FGF23 is also tightly correlated with inflammation in the progression of DN. The aim of this study was to explore whether the C-terminal of FGF23 (FGF23), an antagonist that can block the FGF23 signaling pathway by competing with intact FGF23, could exhibit a therapeutic effect on DN. Read More

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Comparison of solvate ionic liquids and DMSO as an in vivo delivery and storage media for small molecular therapeutics.

BMC Biotechnol 2018 May 29;18(1):32. Epub 2018 May 29.

Metabolic Genetic Diseases Laboratory, Metabolic Research Unit, Deakin University School of Medicine, 75 Pigdons Road, Geelong, VIC, 3216, Australia.

Background: Solvate ionic liquids (SILs) are a new class of ionic liquids that are equimolar solutions of lithium bistrifluoromethanesulfonimide in either triglyme or tetraglyme, referred to as G3LiTFSA and G4LiTFSA, respectively. SILs play a role in energy storage lithium batteries, and have been proposed as potential alternatives to traditional organic solvents such as DMSO. G3TFSA and G4TFSA have been shown to exhibit no toxicity in vivo up to 0. Read More

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May 2018
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The production of a recombinant tandem single chain fragment variable capable of binding prolamins triggering celiac disease.

BMC Biotechnol 2018 May 29;18(1):30. Epub 2018 May 29.

Research Division Biochemical Engineering, Institute of Chemical, Environmental and Bioscience Engineering, TU Wien, Vienna, Austria.

Background: Celiac disease (CD) is one of the most common food-related chronic disorders. It is mediated by the dietary consumption of prolamins, which are storage proteins of different grains. So far, no therapy exists and patients are bound to maintain a lifelong diet to avoid symptoms and long-term complications. Read More

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Characterization of a novel thermostable and xylose-tolerant GH 39 β-xylosidase from Dictyoglomus thermophilum.

BMC Biotechnol 2018 May 21;18(1):29. Epub 2018 May 21.

International Centre for Bamboo and Rattan, 8 Fu Tong East Street, Beijing, 100714, China.

Background: β-D-xylosidase is a vital exoglycosidase with the ability to hydrolyze xylooligosaccharides to xylose and to biotransform some saponins by cleaving outer β-xylose. β-D-xylosidase is widely used as one of the xylanolytic enzymes in a diverse range of applications, such as fuel, food and the pharmaceutical industry; therefore, more and more studies have focused on the thermostable and xylose-tolerant β-D-xylosidases.

Results: A thermostable β-xylosidase gene (xln-DT) of 1509 bp was cloned from Dictyoglomus thermophilum and expressed in E. Read More

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UPF1 silenced cellular model systems for screening of read-through agents active on β39 thalassemia point mutation.

BMC Biotechnol 2018 May 15;18(1):28. Epub 2018 May 15.

Department of Life Sciences and Biotechnology, Biochemistry and Molecular Biology Section, University of Ferrara, Via Fossato di Mortara 74, 44121, Ferrara, Italy.

Background: Nonsense mutations promote premature translational termination, introducing stop codons within the coding region of mRNAs and causing inherited diseases, including thalassemia. For instance, in β39 thalassemia the CAG (glutamine) codon is mutated to the UAG stop codon, leading to premature translation termination and to mRNA destabilization through the well described NMD (nonsense-mediated mRNA decay). In order to develop an approach facilitating translation and, therefore, protection from NMD, ribosomal read-through molecules, such as aminoglycoside antibiotics, have been tested on mRNAs carrying premature stop codons. Read More

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Engineering a vitamin B high-throughput screening system by riboswitch sensor in Sinorhizobium meliloti.

BMC Biotechnol 2018 May 11;18(1):27. Epub 2018 May 11.

Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 32 West 7th Avenue, Tianjin Airport Economic Area, Tianjin, 300308, China.

Background: As a very important coenzyme in the cell metabolism, Vitamin B (cobalamin, VB) has been widely used in food and medicine fields. The complete biosynthesis of VB requires approximately 30 genes, but overexpression of these genes did not result in expected increase of VB production. High-yield VB-producing strains are usually obtained by mutagenesis treatments, thus developing an efficient screening approach is urgently needed. Read More

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Cloning and characterization of enoate reductase with high β-ionone to dihydro-β-ionone bioconversion productivity.

BMC Biotechnol 2018 May 9;18(1):26. Epub 2018 May 9.

International centre for bamboo and rattan, 8 FuTong East Street, Beijing, 100714, China.

Background: Dihydro-β-ionone is a principal aroma compound and has received considerable attention by flavor and fragrance industry. The traditional method of preparing dihydro-β-ionone has many drawbacks, which has restricted its industrial application. Therefore, it is necessary to find a biotechnological method to produce dihydro-β-ionone. Read More

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Effective preparation of a monoclonal antibody against human chromogranin A for immunohistochemical diagnosis.

BMC Biotechnol 2018 May 4;18(1):25. Epub 2018 May 4.

Key Laboratory of Pathogenic Fungi and Mycotoxins of Fujian Province, Key Laboratory of Biopesticide and Chemical Biology of Education Ministry, and School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002, China.

Background: Human chromogranin A (CgA) is a ~ 49 kDa secreted protein mainly from neuroendocrine cells and endocrine cells. The CgA values in the diagnosis of tumor, and in the potential role in prognostic and predictive tumor as a biomarker.

Results: The synthesized gene of CgA coding area was cloned and expressed as fusion protein CgA-His in procaryotic system. Read More

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Development of a baculovirus vector carrying a small hairpin RNA for suppression of sf-caspase-1 expression and improvement of recombinant protein production.

BMC Biotechnol 2018 May 2;18(1):24. Epub 2018 May 2.

College of Life Sciences, Northwest A & F University, Yangling, Shaanxi, 712100, People's Republic of China.

Background: The Baculovirus expression vector system (BEVS) is a transient expression platform for recombinant protein production in insect cells. Baculovirus infection of insect cells will shutoff host translation and induce apoptosis and lead to the termination of protein expression. Previous reports have demonstrated the enhancement of protein yield in BEVS using stable insect cell lines expressing interference RNA to suppress the expression of caspase-1. Read More

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Heavy ion mutagenesis combined with triclosan screening provides a new strategy for improving the arachidonic acid yield in Mortierella alpina.

BMC Biotechnol 2018 May 2;18(1):23. Epub 2018 May 2.

Shandong Provincial Key Laboratory of Energy Genetics, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, 266101, Shandong, China.

Background: Arachidonic acid (ARA), which is a ω-6 polyunsaturated fatty acid, has a wide range of biological activities and is an essential component of cellular membranes in some human tissues. Mortierella alpina is the best strain for industrial production of ARA. To increase its yield of arachidonic acid, heavy ion beam irradiation mutagenesis of Mortierella alpina was carried out in combination with triclosan and octyl gallate treatment. Read More

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Antiplasmodial and trypanocidal activity of violacein and deoxyviolacein produced from synthetic operons.

BMC Biotechnol 2018 Apr 11;18(1):22. Epub 2018 Apr 11.

Cambridge Systems Biology Centre and Department of Biochemistry, University of Cambridge, Cambridge, UK.

Background: Violacein is a deep violet compound that is produced by a number of bacterial species. It is synthesized from tryptophan by a pathway that involves the sequential action of 5 different enzymes (encoded by genes vioA to vioE). Violacein has antibacterial, antiparasitic, and antiviral activities, and also has the potential of inducing apoptosis in certain cancer cells. Read More

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April 2018
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A factor XIa-activatable hirudin-albumin fusion protein reduces thrombosis in mice without promoting blood loss.

BMC Biotechnol 2018 Apr 5;18(1):21. Epub 2018 Apr 5.

Centre for Innovation, Canadian Blood Services, Hamilton, ON, Canada.

Background: Hirudin is a potent thrombin inhibitor but its antithrombotic properties are offset by bleeding side-effects. Because hirudin's N-terminus must engage thrombin's active site for effective inhibition, fusing a cleavable peptide at this site may improve hirudin's risk/benefit ratio as a therapeutic agent. Previously we engineered a plasmin cleavage site (C) between human serum albumin (HSA) and hirudin variant 3 (HV3) in fusion protein HSACHV3. Read More

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April 2018
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Soluble expression of an amebic cysteine protease in the cytoplasm of Escherichia coli SHuffle Express cells and purification of active enzyme.

BMC Biotechnol 2018 Apr 3;18(1):20. Epub 2018 Apr 3.

Facultad de Ciencias Químicas e Ingeniería, Universidad Autónoma de Baja California, Calzada Universidad 14418, Parque Industrial Internacional, 22390, Tijuana, BCN, México.

Background: Recombinant production of amebic cysteine proteases using Escherichia coli cells as the bacterial system has become a challenging effort, with protein insolubility being the most common issue. Since many of these enzymes need a native conformation stabilized by disulfide bonds, an elaborate process of oxidative folding is usually demanded to get a functional protein. The cytoplasm of E. Read More

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April 2018
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Successful production of genome-edited rats by the rGONAD method.

BMC Biotechnol 2018 Apr 2;18(1):19. Epub 2018 Apr 2.

Division of Molecular Genetics, Shigei Medical Research Institute, 2117 Yamada, Minami-ku, Okayama, 701-0202, Japan.

Background: Recent progress in development of the CRISPR/Cas9 system has been shown to be an efficient gene-editing technology in various organisms. We recently developed a novel method called Genome-editing via Oviductal Nucleic Acids Delivery (GONAD) in mice; a novel in vivo genome editing system that does not require ex vivo handling of embryos, and this technology is newly developed and renamed as "improved GONAD" (i-GONAD). However, this technology has been limited only to mice. Read More

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April 2018
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High yield production and purification of two recombinant thermostable phosphotriesterase-like lactonases from Sulfolobus acidocaldarius and Sulfolobus solfataricus useful as bioremediation tools and bioscavengers.

BMC Biotechnol 2018 Mar 20;18(1):18. Epub 2018 Mar 20.

Department of Experimental Medicine, Section of Biotechnology and Molecular Biology, University of Campania "Luigi Vanvitelli"-ex Second University of Naples, Naples, Italy.

Background: Thermostable phosphotriesterase-like lactonases (PLLs) are able to degrade organophosphates and could be potentially employed as bioremediation tools and bioscavengers. But nowadays their manufacturing in high yields is still an issue that limits their industrial applications. In this work we aimed to set up a high yield production and purification biotechnological process of two recombinant PLLs expressed in E. Read More

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March 2018
3 Reads

Cell-based reference samples designed with specific differences in microRNA biomarkers.

BMC Biotechnol 2018 Mar 20;18(1):17. Epub 2018 Mar 20.

Joint Initiative for Metrology in Biology, National Institute of Standards and Technology, 443 Via Ortega, Stanford, CA, 94305, USA.

Background: We demonstrate the feasibility of creating a pair of reference samples to be used as surrogates for clinical samples measured in either a research or clinical laboratory setting. The reference sample paradigm presented and evaluated here is designed to assess the capability of a measurement process to detect true differences between two biological samples. Cell-based reference samples can be created with a biomarker signature pattern designed in silico. Read More

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March 2018
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Total RNA extraction from tissues for microRNA and target gene expression analysis: not all kits are created equal.

BMC Biotechnol 2018 Mar 16;18(1):16. Epub 2018 Mar 16.

Laboratory for Cancer Medicine, Harry Perkins Institute of Medical Research, University of Western Australia, Centre for Medical Research, QEII Medical Centre, 6 Verdun St, Nedlands, WA, 6009, Australia.

Background: microRNAs (miRNAs) are short non-coding RNAs that fine-tune gene expression. The aberrant expression of miRNAs is associated with many diseases and they have both therapeutic and biomarker potential. However, our understanding of their usefulness is dependent on the tools we have to study them. Read More

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March 2018
6 Reads

Phenotype-specific recombinant haptoglobin polymers co-expressed with C1r-like protein as optimized hemoglobin-binding therapeutics.

BMC Biotechnol 2018 Mar 15;18(1):15. Epub 2018 Mar 15.

Division of Internal Medicine, University Hospital Zurich, Ramistrasse 100, CH-8091, Zurich, Switzerland.

Background: Preclinical studies have evaluated haptoglobin (Hp) polymers from pooled human plasma as a therapeutic protein to attenuate toxic effects of cell-free hemoglobin (Hb). Proof of concept studies have demonstrated efficacy of Hp in hemolysis associated with transfusion and sickle cell anemia. However, phenotype-specific Hp products might be desirable to exploit phenotype specific activities of Hp 1-1 versus Hp 2-2, offering opportunities for recombinant therapeutics. Read More

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March 2018
5 Reads

Maintenance of human chondrogenic phenotype on a dendrimer-immobilized surface for an application of cell sheet engineering.

BMC Biotechnol 2018 Mar 14;18(1):14. Epub 2018 Mar 14.

Biological Engineering Program, Faculty of Engineering, King Mongkut's University of Technology Thonburi, Bangkok, 10140, Thailand.

Background: Dedifferentiation of chondrocytes during cell expansion is one of the barriers in tissue construction for cartilage repair. To understand chondrocyte behavior and improve cell expansion in monolayer culture, this study investigated the effects of morphological changes and cellular aggregation on the maintenance of chondrogenic capacity by observing the expression patterns of chondrogenic (collagen type II and aggrecan) and dedifferentiation (collagen type I) markers. Primary human chondrocytes were cultured on either a polystyrene surface (PS) or a polyamidoamine dendrimer surface with a fifth-generation (G5) dendron structure to create a one-step process of cell expansion and the maintenance of chondrogenic activities prior to the construction of cell sheets. Read More

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March 2018
3 Reads

Microencapsulation extends mycelial viability of Streptomyces lividans 66 and increases enzyme production.

BMC Biotechnol 2018 Mar 12;18(1):13. Epub 2018 Mar 12.

Microbial Biotechnology, Institute of Biology Leiden, Leiden University, Sylviusweg 72, 2333, BE, Leiden, The Netherlands.

Background: Filamentous bacteria of the genus Streptomyces produce a large arsenal of industrially relevant antibiotics and enzymes. The industrial production of these molecules occurs in large fermenters, where many streptomycetes form dense mycelial networks called pellets. Pellets are characterized by slow growth and inefficient nutrient transfer and therefore regarded as undesirable from the perspective of productivity. Read More

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March 2018
3 Reads

Novel hemostatic biomolecules based on elastin-like polypeptides and the self-assembling peptide RADA-16.

BMC Biotechnol 2018 Mar 7;18(1):12. Epub 2018 Mar 7.

Lab of Tissue Engineering, Provincial Key Laboratory of Biotechnology of Shaanxi, College of Life Sciences, Northwest University, Taibai North Rd 229, Xi'an, Shaanxi Province, 710069, China.

Background: Safe and effective hemostatic materials are important for reducing mortality resulting from excessive hemorrhage. In this work, new biomaterials with hemostatic effects were created by fusing the gene coding for RADA-16, a self-assembling peptide with the sequence RADARADARADARADA, to the 3'-end of the open reading frame (ORF) encoding elastin-like polypeptides through gene recombination.

Results: The fusion proteins, termed 36R, 60R and 96R, were solubly over-expressed in Escherichia coli BL21 (DE3) based on genetic manipulation of the high-efficiency prokaryotic expression vector pET28a (+) and bacterial transformation. Read More

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March 2018
3 Reads

Agrobacterium-mediated and electroporation-mediated transformation of Chlamydomonas reinhardtii: a comparative study.

BMC Biotechnol 2018 Feb 17;18(1):11. Epub 2018 Feb 17.

ENEA, Italian National Agency for New Technologies, Energy and Sustainable Economic Development, Casaccia Research Center, 00123, Rome, Italy.

Background: Chlamydomonas reinhardtii is an unicellular green alga used for functional genomics studies and heterologous protein expression. A major hindrance in these studies is the low level and instability of expression of nuclear transgenes, due to their rearrangement and/or silencing over time.

Results: We constructed dedicated vectors for Agrobacterium-mediated transformation carrying, within the T-DNA borders, the Paromomycin (Paro) selectable marker and an expression cassette containing the Luciferase (Luc) reporter gene. Read More

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February 2018
1 Read

Effects of substrate binding site residue substitutions of xynA from Bacillus amyloliquefaciens on substrate specificity.

BMC Biotechnol 2018 Feb 13;18(1). Epub 2018 Feb 13.

P. G. Department of Biosciences, UGC-Centre of advanced studies, Satellite campus, Sardar Patel University, Sardar Patel Maidan, Bakrol-Vadtal Road, PO Box 39, Vallabh Vidyanagar, Gujarat, 388 120, India.

Background: The aromatic residues of xylanase enzyme, W187, Y124, W144, Y128 and W63 of substrate binding pocket from Bacillus amyloliquefaciens were investigated for their role in substrate binding by homology modelling and sequence analysis. These residues are highly conserved and play an important role in substrate binding through steric hindrance. The substitution of these residues with alanine allows the enzyme to accommodate nonspecific substrates. Read More

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February 2018
2 Reads

Genetically encoded calcium indicator with NTnC-like design and enhanced fluorescence contrast and kinetics.

BMC Biotechnol 2018 Feb 13;18(1):10. Epub 2018 Feb 13.

Moscow Institute of Physics and Technology, Moscow, 123182, Russia.

Background: The recently developed genetically encoded calcium indicator (GECI), called NTnC, has a novel design with reduced size due to utilization of the troponin C (TnC) as a Ca-binding moiety inserted into the mNeonGreen fluorescent protein. NTnC binds two times less Ca ions while maintaining a higher fluorescence brightness at the basal level of Ca in neurons as compared with the calmodulin-based GECIs, such as GCaMPs. In spite of NTnC's high brightness, pH-stability, and high sensitivity to single action potentials, it has a limited fluorescence contrast (F/F) and slow Ca dissociation kinetics. Read More

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February 2018
5 Reads

A new approach for detecting adventitious viruses shows Sf-rhabdovirus-negative Sf-RVN cells are suitable for safe biologicals production.

BMC Biotechnol 2018 Feb 7;18(1). Epub 2018 Feb 7.

GlycoBac LLC, 1938 Harney Street, Laramie, WY, 82072, USA.

Background: Adventitious viral contamination in cell substrates used for biologicals production is a major safety concern. A powerful new approach that can be used to identify adventitious viruses is a combination of bioinformatics tools with massively parallel sequencing technology. Typically, this involves mapping or BLASTN searching individual reads against viral nucleotide databases. Read More

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February 2018
2 Reads

DNA methylome profiling at single-base resolution through bisulfite sequencing of 5mC-immunoprecipitated DNA.

BMC Biotechnol 2018 Feb 6;18(1). Epub 2018 Feb 6.

Department of Stomatology, Chinese PLA General Hospital, Beijing, 100853, People's Republic of China.

Background: Detection of DNA methylome at single-base resolution is a significant challenge but promises to shed considerable light on human disease etiology. Current technologies could not detect DNA methylation genome-wide at single-base resolution with small amount of sequencing data and could not avoid detecting the methylation of repetitive elements which are considered as "junk DNA".

Methods: In this study, we have developed a novel DNA methylome profiling technology named MB-seq with its ability to identify genome-wide 5mC and quantify DNA methylation levels by introduced an assistant adapter AluI-linker This linker can be ligated to sonicated DNA and then be digested after the bisulfite treatment and amplification, which has no effect of MeDIP enrichment. Read More

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February 2018
4 Reads

Two approaches for estimating the lower limit of quantitation (LLOQ) of microRNA levels assayed as exploratory biomarkers by RT-qPCR.

BMC Biotechnol 2018 Feb 2;18(1). Epub 2018 Feb 2.

Center for Drug Evaluation and Research, Food and Drug Administration, Silver Spring, MD, 20993, USA.

Background: Circulating microRNAs are undergoing exploratory use as safety biomarkers in drug development. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) is one common approach used to quantitate levels of microRNAs in samples that includes the use of a standard curve of calibrators fit to a regression model. Guidelines are needed for setting assay quantitation thresholds that are appropriate for this method and to biomarker pre-validation. Read More

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February 2018
9 Reads

Genetic engineering of Escherichia coli to improve L-phenylalanine production.

BMC Biotechnol 2018 Jan 30;18(1). Epub 2018 Jan 30.

Tianjin Institutes of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, People's Republic of China.

Background: L-phenylalanine (L-Phe) is an essential amino acid for mammals and applications expand into human health and nutritional products. In this study, a system level engineering was conducted to enhance L-Phe biosynthesis in Escherichia coli.

Results: We inactivated the PTS system and recruited glucose uptake via combinatorial modulation of galP and glk to increase PEP supply in the Xllp01 strain. Read More

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January 2018
2 Reads

Optimized DNA electroporation for primary human T cell engineering.

BMC Biotechnol 2018 Jan 30;18(1). Epub 2018 Jan 30.

Laboratory of Vaccine and Antibody Engineering, Beijing Institute of Biotechnology, No. 20, Dongdajie street, Fengtai District, Beijing, 100071, China.

Background: Effective gene-delivery systems for primary human T cell engineering are useful tools for both basic research and clinical immunotherapy applications. Pseudovirus-based systems and electro-transfection are the most popular strategies for genetic material transduction. Compared with viral-particle-mediated approaches, electro-transfection is theoretically safer, because it does not promote transgene integration into the host genome. Read More

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January 2018
2 Reads

RNase I -treated quantitative PCR for dsRNA quantitation of RNAi trait in genetically modified crops.

BMC Biotechnol 2018 Jan 17;18(1). Epub 2018 Jan 17.

Dow AgroSciences LLC, 9330 Zionsville Rd, Indianapolis, IN, 46268, USA.

Background: RNA interference (RNAi) technology has been widely used to knockdown target genes via post-transcriptional silencing. In plants, RNAi is used as an effective tool with diverse applications being developed such as resistance against insects, fungi, viruses, and metabolism manipulation. To develop genetically modified (GM) RNAi traits for insect control, a transgene is created and composed of an inversely-repeated sequence of the target gene with a spacer region inserted between the repeats. Read More

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January 2018
2 Reads

Donor age affects proteome composition of tenocyte-derived engineered tendon.

BMC Biotechnol 2018 Jan 16;18(1). Epub 2018 Jan 16.

Institute of Ageing and Chronic Disease, University of Liverpool, William Duncan Building, 6 West Derby Street, Liverpool, L7 8TX, UK.

Background: The concept of tissue engineering is to deliver to the injury site biological scaffolds carrying functional cells that will enhance healing response. The preferred cell source is autologous in order to reduce immune response in the treated individual. However, in elderly patients age-related changes in synthetic activity of the implanted cells and subsequent alterations in tissue protein content may affect therapeutic outcomes. Read More

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January 2018
2 Reads

The function of tcf3 in medaka embryos: efficient knockdown with pePNAs.

BMC Biotechnol 2018 Jan 9;18(1). Epub 2018 Jan 9.

Department for Applied Life Sciences, University of Applied Sciences, FH Campus Wien, Helmut-Qualtinger-Gasse 2, A-1030, Vienna, Austria.

Background: The application of antisense molecules, such as morpholino oligonucleotides, is an efficient method of gene inactivation in vivo. We recently introduced phosphonic ester modified peptide nucleic acids (PNA) for in vivo loss-of-function experiments in medaka embryos. Here we tested novel modifications of the PNA backbone to knockdown the medaka tcf3 gene. Read More

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January 2018
2 Reads

Laser-fabricated cell patterning stencil for single cell analysis.

BMC Biotechnol 2017 Dec 19;17(1):89. Epub 2017 Dec 19.

Center for Biosignatures Discovery Automation, The Biodesign Institute, Arizona State University, 1001 S. McAllister Ave., P.O. Box 877101, Tempe, AZ, 85287-7101, USA.

Precise spatial positioning and isolation of mammalian cells is a critical component of many single cell experimental methods and biological engineering applications. Although a variety of cell patterning methods have been demonstrated, many of these methods subject cells to high stress environments, discriminate against certain phenotypes, or are a challenge to implement. Here, we demonstrate a rapid, simple, indiscriminate, and minimally perturbing cell patterning method using a laser fabricated polymer stencil. Read More

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December 2017
5 Reads

Computer face-matching technology using two-dimensional photographs accurately matches the facial gestalt of unrelated individuals with the same syndromic form of intellectual disability.

BMC Biotechnol 2017 Dec 19;17(1):90. Epub 2017 Dec 19.

Imagus Technology, Brisbane, QLD, Australia.

Background: Massively parallel genetic sequencing allows rapid testing of known intellectual disability (ID) genes. However, the discovery of novel syndromic ID genes requires molecular confirmation in at least a second or a cluster of individuals with an overlapping phenotype or similar facial gestalt. Using computer face-matching technology we report an automated approach to matching the faces of non-identical individuals with the same genetic syndrome within a database of 3681 images [1600 images of one of 10 genetic syndrome subgroups together with 2081 control images]. Read More

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December 2017
3 Reads

Modulation of the regioselectivity of Thermomyces lanuginosus lipase via biocatalyst engineering for the Ethanolysis of oil in fully anhydrous medium.

BMC Biotechnol 2017 Dec 16;17(1):88. Epub 2017 Dec 16.

Department of Biocatalysis, Institute of Catalysis and Petrochemistry (ICP) CSIC. Campus UAM, Cantoblanco, 28049, Madrid, Spain.

Background: Enzymatic ethanolysis of oils (for example, high oleic sunflower oil containing 90% of oleic acid) may yield two different reaction products depending on the regioselectivity of the immobilized lipase biocatalyst. Some lipase biocatalysts exhibit a 1,3-regioselectivity and they produced 2 mols of fatty acid ethyl ester plus 1 mol of sn2-monoacylglycerol (2-MAG) per mol of triglyceride without the release of glycerol. Other lipase biocatalysts are completely non-regioselective releasing 3 mols of fatty acid ethyl ester and 1 mol of glycerol per mol of triglyceride. Read More

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December 2017
4 Reads

Heterologous expression, purification and function of the extracellular domain of human RANK.

BMC Biotechnol 2017 Dec 4;17(1):87. Epub 2017 Dec 4.

Key Laboratory of Stem Cell Biology, Shanghai Institutes for Biological Sciences, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai, 200031, China.

Background: Receptor activator of NF-κB ligand (RANKL)/RANK signaling essentially functions within the skeletal system, particularly participating in osteoclastogenesis and bone resorption. In addition, this signaling pathway has also been shown to influence tumor progression as well as the development and function of the immune system. Therefore, blocking the interaction between RANKL and RANK is a new therapeutic approach to prevent bone-related diseases and cancer. Read More

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December 2017
6 Reads

A tandem CBM25 domain of α-amylase from Microbacterium aurum as potential tool for targeting proteins to starch granules during starch biosynthesis.

BMC Biotechnol 2017 Dec 4;17(1):86. Epub 2017 Dec 4.

Wageningen University and Research, Plant Breeding, P.O. Box 386, 6700, AJ, Wageningen, The Netherlands.

Background: Starch-binding domains from carbohydrate binding module family 20 have been used as a tool for starch engineering. Previous studies showed that expression of starch binding domain fusion proteins in planta resulted in modified starch granule structures and physicochemical properties. However, although 13 carbohydrate binding module families have been reported to contain starch-binding domains, only starch-binding domains from carbohydrate binding module family 20 have been well studied and introduced into plants successfully. Read More

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December 2017
8 Reads

Optimal reduction of chemical oxygen demand and NH-N from landfill leachate using a strongly resistant novel Bacillus salmalaya strain.

BMC Biotechnol 2017 Nov 28;17(1):85. Epub 2017 Nov 28.

Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia.

Background: When the unavoidable waste generation is considered as damaging to our environment, it becomes crucial to develop a sustainable technology to remediate the pollutant source towards an environmental protection and safety. The development of a bioengineering technology for highly efficient pollutant removal is this regard. Given the high ammonia nitrogen content and chemical oxygen demand of landfill leachate, Bacillus salmalaya strain 139SI, a novel resident strain microbe that can survive in high ammonia nitrogen concentrations, was investigated for the bioremoval of ammonia nitrogen from landfill leachate. Read More

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November 2017
5 Reads

Characterization of a novel bioflocculant from a marine bacterium and its application in dye wastewater treatment.

BMC Biotechnol 2017 Nov 17;17(1):84. Epub 2017 Nov 17.

Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, People's Republic of China.

Background: The identification of microorganisms with excellent flocculant-producing capability and optimization of the fermentation process are necessary for the wide-scale application of bioflocculants. Thus, we evaluated the flocculant-producing ability of a novel strain identified by the screening of marine bacteria, and we report for the first time the properties of the bioflocculant produced by Alteromonas sp. in the treatment of dye wastewater. Read More

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November 2017
9 Reads

A new single-step protocol for rapid baculovirus-driven protein production in insect cells.

BMC Biotechnol 2017 Nov 16;17(1):83. Epub 2017 Nov 16.

Max-Planck Institute of Biochemistry, Am Klopferspitz 18, 82152, Martinsried, Germany.

Background: In the last three decades, the Baculovirus expression vector system (BEV) has evolved to one of the most widely used eukaryotic systems for heterologous protein expression including approved vaccines and therapies. Despite the significant improvements introduced during the past years, the BEV system still has major drawbacks, primarily the time required to generate recombinant virus and virus instability for certain target proteins. In this study we show that the conventional method to generate recombinant Baculovirus using a Tn7 transposition based system can be shortened to a single-step transfection-only procedure without further amplification. Read More

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November 2017
5 Reads
2.03 Impact Factor

Comparison of [HSO], [Cl] and [MeCO] as anions in pretreatment of aspen and spruce with imidazolium-based ionic liquids.

BMC Biotechnol 2017 Nov 15;17(1):82. Epub 2017 Nov 15.

Department of Chemistry, Umeå University, SE-901 87, Umeå, Sweden.

Background: Ionic liquids (ILs) draw attention as green solvents for pretreatment of lignocellulose before enzymatic saccharification. Imidazolium-based ILs with different anionic constituents ([HSO], [Cl], [MeCO]) were compared with regard to pretreatment of wood from aspen and spruce. The objective was to elucidate how the choice of anionic constituent affected the suitability of using the IL for pretreatment of hardwood, such as aspen, and softwood, such as spruce. Read More

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November 2017
5 Reads

AFEAP cloning: a precise and efficient method for large DNA sequence assembly.

BMC Biotechnol 2017 Nov 14;17(1):81. Epub 2017 Nov 14.

McGovern Medical School, the University of Texas Health Science Center at Houston, Houston, 77030, USA.

Background: Recent development of DNA assembly technologies has spurred myriad advances in synthetic biology, but new tools are always required for complicated scenarios. Here, we have developed an alternative DNA assembly method named AFEAP cloning (Assembly of Fragment Ends After PCR), which allows scarless, modular, and reliable construction of biological pathways and circuits from basic genetic parts.

Methods: The AFEAP method requires two-round of PCRs followed by ligation of the sticky ends of DNA fragments. Read More

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November 2017
5 Reads

Adaptation of the GoldenBraid modular cloning system and creation of a toolkit for the expression of heterologous proteins in yeast mitochondria.

BMC Biotechnol 2017 Nov 13;17(1):80. Epub 2017 Nov 13.

Centro de Biotecnología y Genómica de Plantas, Universidad Politécnica de Madrid (UPM) - Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Campus Montegancedo UPM, 28223, Pozuelo de Alarcón, Madrid, Spain.

Background: There is a need for the development of synthetic biology methods and tools to facilitate rapid and efficient engineering of yeast that accommodates the needs of specific biotechnology projects. In particular, the manipulation of the mitochondrial proteome has interesting potential applications due to its compartmentalized nature. One of these advantages resides in the fact that metalation occurs after protein import into mitochondria, which contains pools of iron, zinc, copper and manganese ions that can be utilized in recombinant metalloprotein metalation reactions. Read More

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November 2017
6 Reads