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BMC Biotechnology

Adrián Montaño, Maribel Forero-Castro, Jesús-María Hernández-Rivas, Ignacio García-Tuñón, Rocío Benito
BACKGROUND: Genome editing technologies offers new opportunities for tackling diseases such as acute lymphoblastic leukemia (ALL) that have been beyond the reach of previous therapies. RESULTS: We show how the recent availability of genome-editing tools such as CRISPR-Cas9 are an important means of advancing functional studies of ALL through the incorporation, elimination and modification of somatic mutations and fusion genes in cell lines and mouse models. These tools not only broaden the understanding of the involvement of various genetic alterations in the pathogenesis of the disease but also identify new therapeutic targets for future clinical trials...
July 17, 2018: BMC Biotechnology
Qin Liu, Hui-Jie Dang, Yuan-Hang Wu, Min Li, Yin-Hua Chen, Xiao-Lei Niu, Kai-Mian Li, Li-Juan Luo
BACKGROUND: With the rapid development of sequencing technologies, increasing amount of genomic information has been accumulated. To clone genes for further functional studies in large scale, a cheap, fast and efficient cloning vector is desired. RESULTS: A bifunctional vector pXST has been constructed. The pXST vector harbors a XcmI-ccdB-XcmI cassette and restriction site SmaI. Digestion the vector with XcmI generates a single thymidine (T) overhang at 3' end which facilitates TA cloning, and SmaI gives blunt end that enables the blunt-end ligation...
July 13, 2018: BMC Biotechnology
Zi-Lu Liu, Hua-Nan Li, Hui-Ting Song, Wen-Jing Xiao, Wu-Cheng Xia, Xiao-Peng Liu, Zheng-Bing Jiang
BACKGROUND: Cellulose is the most important component of lignocellulose, and its degradation requires three different types of enzymes to act synergistically. There have been reports of single gene duality, but no gene has been described to have more than two functions. Cloning and expression of fusion cellulases containing more than two kinds of catalytic domains has not been reported thus far. RESULTS: We synthesized three different cellulase genes and linked the three catalytic domains with a (G4 S)3 flexible linker...
July 13, 2018: BMC Biotechnology
Rashmi Sharma, Fetweh H Al-Saleem, Rama Devudu Puligedda, Amy Rattelle, David R Lynch, Scott K Dessain
BACKGROUND: Anti-NMDA receptor encephalitis (ANRE) is a potentially lethal disease attributed to auto-antibodies against the N-methyl-D-aspartate receptor (NMDAR). Full recovery is possible if therapy is initiated early in the disease course. Detection of ANRE antibodies in the cerebrospinal fluid (CSF) is essential for diagnosis. The assays for ANRE-associated IgGs often rely on cells transiently transfected with NMDAR genes. A cell line that stably expresses pathogenic NMDAR epitopes could improve standardization of the assays and provide antigen that could be used in commercial solid state assay systems...
June 27, 2018: BMC Biotechnology
Martinus J A Daas, Bart Nijsse, Antonius H P van de Weijer, Bart W A J Groenendaal, Fons Janssen, John van der Oost, Richard van Kranenburg
BACKGROUND: Consolidated bioprocessing (CBP) is a cost-effective approach for the conversion of lignocellulosic biomass to biofuels and biochemicals. The enzymatic conversion of cellulose to glucose requires the synergistic action of three types of enzymes: exoglucanases, endoglucanases and β-glucosidases. The thermophilic, hemicellulolytic Geobacillus thermodenitrificans T12 was shown to harbor desired features for CBP, although it lacks the desired endo and exoglucanases required for the conversion of cellulose...
June 27, 2018: BMC Biotechnology
Marie-Alice Fraiture, Julie Vandamme, Philippe Herman, Nancy H C Roosens
BACKGROUND: Recently, an integrated DNA walking strategy has been proposed to prove the presence of GMO via the characterisation of sequences of interest, including their transgene flanking regions and the unnatural associations of elements in their transgenic cassettes. To this end, the p35S, tNOS and t35S pCAMBIA elements have been selected as key targets, allowing the coverage of most of GMO, EU authorized or not. In the present study, a bidirectional DNA walking method anchored on the CryAb/c genes is proposed with the aim to cover additional GMO and additional sequences of interest...
June 27, 2018: BMC Biotechnology
Yasuhiro Watanabe, Takashi Matsuba, Mami Nakanishi, Mio Une, Ritsuko Hanajima, Kenji Nakashima
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. In this study, we used Bcl-2, a protein known to have anti-apoptotic properties in vivo and in vitro, as a parcel to couple to TeNT fragments...
June 11, 2018: BMC Biotechnology
Yuliya Khilko, Philip D Weyman, John I Glass, Mark D Adams, Melanie A McNeil, Peter B Griffin
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. The purpose of this study was to develop a gene assembly protocol utilizing a digital microfluidic device...
June 1, 2018: BMC Biotechnology
Pengfei Zhang, Xianghua Yuan, Yuguang Du, Jian-Jun Li
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. The use of P. pastoris for the expression of recombinant proteins has become popular. The endoglucanase from Reticulitermes speratus (RsEG), belonging to glycoside hydrolase family 9 (GHF9), has not been produced in P...
June 1, 2018: BMC Biotechnology
Hong-Yan Zhao, Hong Feng
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. The alkaline serine protease (DHAP) from Bacillus pumilus BA06 is a typical mesophilic enzyme, which has demonstrated great potential in various industrial applications...
June 1, 2018: BMC Biotechnology
Jean-Michel Michno, Robert M Stupar
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. Analytical pipelines that do not apply proper thresholds nor assess reproducibility among samples are susceptible to calling false-positive variants...
June 1, 2018: BMC Biotechnology
Nanyu Han, Huabiao Miao, Tingting Yu, Bo Xu, Yunjuan Yang, Qian Wu, Rui Zhang, Zunxi Huang
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...
June 1, 2018: BMC Biotechnology
Xiaomin Zhang, Kaiwen Guo, Feng Xia, Xinyu Zhao, Zhifeng Huang, Jianlou Niu
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 (FGF23C-tail ), an antagonist that can block the FGF23 signaling pathway by competing with intact FGF23, could exhibit a therapeutic effect on DN...
May 30, 2018: BMC Biotechnology
Hua Li, Yonghua Xu, Weiming Yang, Xiaohong Huang
BACKGROUND: According to recent studies, the phospholipase A2 receptor 1 (PLA2 R1) may be used as a biomarker to diagnose idiopathic membranous nephropathy (iMN). Moreover, the immune-dominant regions of PLA2 R1 have been identified. The aim of the present study was to construct a diagnostic antigen based on the immune-dominant region of PLA2 R1 and develop a specific serological detection method for PLA2 R1 antibodies. RESULTS: The tandem multi-epitope diagnostic antigen (designated 'R101'), which includes aa 39-130 (CysR), aa 238-356 (CTLD1), and aa 1136-1234 (CTLD7) of PLA2 R1; thioredoxin at the N-terminus; and a His tag at the C-terminus, was prepared at a concentration of 2...
May 29, 2018: BMC Biotechnology
Prusothman Yoganantharajah, Alexander P Ray, Daniel J Eyckens, Luke C Henderson, Yann Gibert
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 0.5% (v/v), and solubilize small compounds (N,N-diethylaminobenzaldehyde) with full penetrance, similar to DMSO delivered DEAB...
May 29, 2018: BMC Biotechnology
Britta Eggenreich, Elke Scholz, David Johannes Wurm, Florian Forster, Oliver Spadiut
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. To support those patients we developed a tandem single chain Fragment variable (tscFv) acting as a neutralizing agent against prolamins. We recombinantly produced this molecule in E...
May 29, 2018: BMC Biotechnology
Qi Li, Tao Wu, Zhipeng Qi, Linguo Zhao, Jianjun Pei, Feng Tang
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...
May 21, 2018: BMC Biotechnology
Francesca Salvatori, Mariangela Pappadà, Giulia Breveglieri, Elisabetta D'Aversa, Alessia Finotti, Ilaria Lampronti, Roberto Gambari, Monica Borgatti
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 β0 39 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...
May 15, 2018: BMC Biotechnology
Yingying Cai, Miaomiao Xia, Huina Dong, Yuan Qian, Tongcun Zhang, Beiwei Zhu, Jinchuan Wu, Dawei Zhang
BACKGROUND: As a very important coenzyme in the cell metabolism, Vitamin B12 (cobalamin, VB12 ) has been widely used in food and medicine fields. The complete biosynthesis of VB12 requires approximately 30 genes, but overexpression of these genes did not result in expected increase of VB12 production. High-yield VB12 -producing strains are usually obtained by mutagenesis treatments, thus developing an efficient screening approach is urgently needed. RESULT: By the help of engineered strains with varied capacities of VB12 production, a riboswitch library was constructed and screened, and the btuB element from Salmonella typhimurium was identified as the best regulatory device...
May 11, 2018: BMC Biotechnology
Xuesong Zhang, Shiyong Liao, Fuliang Cao, Linguo Zhao, Jianjun Pei, Feng Tang
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. RESULTS: In this study, the enoate reductase with high conversion efficiency of β-ionone to dihydro-β-ionone, DBR1, was obtained by screening four genetically engineered bacteria...
May 9, 2018: BMC Biotechnology
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