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BMC Biotechnology

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https://www.readbyqxmd.com/read/30200961/development-of-a-vitrification-method-for-preserving-human-myoblast-cell-sheets-for-myocardial-regeneration-therapy
#1
Hirotatsu Ohkawara, Shigeru Miyagawa, Satsuki Fukushima, Shin Yajima, Atsuhiro Saito, Hiroshi Nagashima, Yoshiki Sawa
BACKGROUND: Tissue-engineered cardiac constructs have potential in the functional recovery of heart failure; however, the preservation of these constructs is crucial for the development and widespread application of this treatment. We hypothesized that tissue-engineered skeletal myoblast (SMB) constructs may be preserved by vitrification to conserve biological function and structure. RESULTS: Scaffold-free cardiac cell-sheet constructs were prepared from SMBs and immersed in a vitrification solution containing ethylene glycol, sucrose, and carboxyl poly-L-lysine...
September 10, 2018: BMC Biotechnology
https://www.readbyqxmd.com/read/30200951/a-novel-synthetic-trivalent-single-chain-variable-fragment-tri-scfv-construction-platform-based-on-the-spytag-spycatcher-protein-ligase-system
#2
Md Kausar Alam, Michelle Brabant, Raja Solomon Viswas, Kris Barreto, Humphrey Fonge, C Ronald Geyer
BACKGROUND: Advances in antibody engineering provide strategies to construct recombinant antibody-like molecules with modified pharmacokinetic properties. Multermerization is one strategy that has been used to produce antibody-like molecules with two or more antigen binding sites. Multimerization enhances the functional affinity (avidity) and can be used to optimize size and pharmacokinetic properties. Most multimerization strategies involve genetically fusing or non-covalently linking antibody fragments using oligomerization domains...
September 10, 2018: BMC Biotechnology
https://www.readbyqxmd.com/read/30180895/overexpression-of-a-rice-bahd-acyltransferase-gene-in-switchgrass-panicum-virgatum-l-enhances-saccharification
#3
Guotian Li, Kyle C Jones, Aymerick Eudes, Venkataramana R Pidatala, Jian Sun, Feng Xu, Chengcheng Zhang, Tong Wei, Rashmi Jain, Devon Birdseye, Patrick E Canlas, Edward E K Baidoo, Phat Q Duong, Manoj K Sharma, Seema Singh, Deling Ruan, Jay D Keasling, Jenny C Mortimer, Dominique Loqué, Laura E Bartley, Henrik V Scheller, Pamela C Ronald
BACKGROUND: Switchgrass (Panicum virgatum L.) is a promising bioenergy feedstock because it can be grown on marginal land and produces abundant biomass. Recalcitrance of the lignocellulosic components of the switchgrass cell wall to enzymatic degradation into simple sugars impedes efficient biofuel production. We previously demonstrated that overexpression of OsAT10, a BAHD acyltransferase gene, enhances saccharification efficiency in rice. RESULTS: Here we show that overexpression of the rice OsAT10 gene in switchgrass decreased the levels of cell wall-bound ferulic acid (FA) in green leaf tissues and to a lesser extent in senesced tissues, and significantly increased levels of cell wall-bound p-coumaric acid (p-CA) in green leaves but decreased its level in senesced tissues of the T0 plants under greenhouse conditions...
September 4, 2018: BMC Biotechnology
https://www.readbyqxmd.com/read/30170595/a-rapid-in-vitro-method-to-flip-back-the-double-floxed-inverted-open-reading-frame-in-a-plasmid
#4
Jian Xu, Yongling Zhu
BACKGROUND: The LoxP site based genetic switch, the FLEX, also known as DIO (Double-Floxed Inverted Open reading frame), was invented to turn on gene expression via Cre-mediated recombination. Since its first publication, numerous FLEX switch plasmids have been generated. These plasmids are designed to only work in combination with Cre. However on many occasions it is necessary to covert these FLEX plasmids back into constitutive expression plasmids so that they can also be used in non-Cre-expressing cells and in non-genetically modified animal models...
August 31, 2018: BMC Biotechnology
https://www.readbyqxmd.com/read/30170578/heterologous-expression-of-cyclodextrin-glycosyltransferase-from-paenibacillus-macerans-in-escherichia-coli-and-its-application-in-2-o-%C3%AE-d-glucopyranosyl-l-ascorbic-acid-production
#5
Yujia Jiang, Jie Zhou, Ruofan Wu, Fengxue Xin, Wenming Zhang, Yan Fang, Jiangfeng Ma, Weiliang Dong, Min Jiang
BACKGROUND: Cyclodextrin glucanotransferase (CGTase) can transform L-ascorbic acid (L-AA, vitamin C) to 2-O-α-D-glucopyranosyl-L-ascorbic acid (AA-2G), which shows diverse applications in food, cosmetic and pharmaceutical industries. RESULTS: In this study, the cgt gene encoding α-CGTase from Paenibacillus macerans was codon-optimized (opt-cgt) and cloned into pET-28a (+) for intracellular expression in E. coli BL21 (DE3). The Opt-CGT was purified by Ni2+ -NTA resin with a 55% recovery, and specific activity was increased significantly from 1...
August 31, 2018: BMC Biotechnology
https://www.readbyqxmd.com/read/30157831/oil-body-bound-oleosin-rhfgf9-fusion-protein-expressed-in-safflower-carthamus-tinctorius-l-stimulates-hair-growth-and-wound-healing-in-mice
#6
Jingbo Cai, Ruicheng Wen, Wenqing Li, Xiuran Wang, Haishan Tian, Shanyong Yi, Linbo Zhang, Xiaokun Li, Chao Jiang, Haiyan Li
BACKGROUND: Fibroblast growth factor 9 (FGF9) is a heparin-binding growth factor, secreted by both mesothelial and epithelial cells, which participates in hair follicle regeneration, wound healing, and bone development. A suitable source of recombinant human FGF9 (rhFGF9) is needed for research into potential clinical applications. We present that expression of oleosin-rhFGF9 fusion protein in safflower (Carthamus tinctorius L.) seeds stimulates hair growth and wound healing. RESULTS: The oleosin-rhFGF9 expressed in safflower seeds, in which it localizes to the surface of oil bodies...
August 29, 2018: BMC Biotechnology
https://www.readbyqxmd.com/read/30134885/a-paratransgenic-strategy-to-block-transmission-of-xylella-fastidiosa-from-the-glassy-winged-sharpshooter-homalodisca-vitripennis
#7
Arinder K Arora, Kendra N Pesko, Verónica Quintero-Hernández, Lourival D Possani, Thomas A Miller, Ravi V Durvasula
BACKGROUND: Arthropod-borne diseases remain a leading cause of human morbidity and mortality and exact an enormous toll on global agriculture. The practice of insecticide-based control is fraught with issues of excessive cost, human and environmental toxicity, unwanted impact on beneficial insects and selection of resistant insects. Efforts to modulate insects to eliminate pathogen transmission have gained some traction and remain future options for disease control. RESULTS: Here, we report a paratransgenic strategy that targets transmission of Xylella fastidiosa, a leading bacterial pathogen of agriculture, by the Glassy-Winged Sharpshooter (GWSS), Homalodisca vitripennis...
August 22, 2018: BMC Biotechnology
https://www.readbyqxmd.com/read/30119626/lignocellulose-integration-to-1g-ethanol-process-using-filamentous-fungi-fermentation-prospects-of-edible-strain-of-neurospora-intermedia
#8
Ramkumar B Nair, Osagie A Osadolor, Vamsi K Ravula, Patrik R Lennartsson, Mohammad J Taherzadeh
BACKGROUND: Integration of first- and second-generation ethanol processes is one among the alternate approaches that efficiently address the current socio-economic issues of the bioethanol sector. Edible filamentous fungus capable of utilizing pentoses from lignocelluloses and also possessing biomass application as potential animal feed component was used as the fermentation strain for the integration model. This study presents various fermentation aspects of using edible filamentous fungi in the integrated first and second generation ethanol process model...
August 17, 2018: BMC Biotechnology
https://www.readbyqxmd.com/read/30092808/acetosyringone-treatment-duration-affects-large-t-dna-molecule-transfer-to-rice-callus
#9
Jing Xi, Minesh Patel, Shujie Dong, Qiudeng Que, Rongda Qu
BACKGROUND: Large T-DNA fragment transfer has long been a problem for Agrobacterium-mediated transformation. Although vector systems, such as the BIBAC series, were successfully developed for the purpose, low transformation efficiencies were consistently observed. RESULTS: To gain insights of this problem in monocot transformation, we investigated the T-strand accumulation of various size of T-DNA in two kinds of binary vectors (one copy vs. multi-copy) upon acetosyringone (AS) induction and explored ways to improve the efficiency of the large T-DNA fragment transfer in Agrobacterium-mediated rice transformation...
August 9, 2018: BMC Biotechnology
https://www.readbyqxmd.com/read/30081895/expression-of-novel-fusion-antiviral-proteins-ricin-a-chain-pokeweed-antiviral-proteins-rta-paps-in-escherichia-coli-and-their-inhibition-of-protein-synthesis-and-of-hepatitis-b-virus-in-vitro
#10
Yasser Hassan, Sherry Ogg, Hui Ge
BACKGROUND: Ricin A chain (RTA) and Pokeweed antiviral proteins (PAPs) are plant-derived N-glycosidase ribosomal-inactivating proteins (RIPs) isolated from Ricinus communis and Phytolacca Americana respectively. This study was to investigate the potential production amenability and sub-toxic antiviral value of novel fusion proteins between RTA and PAPs (RTA-PAPs). In brief, RTA-Pokeweed antiviral protein isoform 1 from seeds (RTA-PAPS1) was produced in an E. coli in vivo expression system, purified from inclusion bodies using gel filtration chromatography and protein synthesis inhibitory activity assayed by comparison to the production of a control protein Luciferase...
August 6, 2018: BMC Biotechnology
https://www.readbyqxmd.com/read/30041697/the-nicotine-degrading-enzyme-nica2-reduces-nicotine-levels-in-blood-nicotine-distribution-to-brain-and-nicotine-discrimination-and-reinforcement-in-rats
#11
Paul R Pentel, Michael D Raleigh, Mark G LeSage, Thomas Thisted, Stephen Horrigan, Zuzana Biesova, Matthew W Kalnik
BACKGROUND: The bacterial nicotine-degrading enzyme NicA2 isolated from P. putida was studied to assess its potential use in the treatment of tobacco dependence. RESULTS: Rats were pretreated with varying i.v. doses of NicA2, followed by i.v. administration of nicotine at 0.03 mg/kg. NicA2 had a rapid onset of action reducing blood and brain nicotine concentrations in a dose-related manner, with a rapid onset of action. A 5 mg/kg NicA2 dose reduced the nicotine concentration in blood by > 90% at 1 min after the nicotine dose, compared to controls...
July 24, 2018: BMC Biotechnology
https://www.readbyqxmd.com/read/30016959/targeted-genome-editing-in-acute-lymphoblastic-leukemia-a-review
#12
REVIEW
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
https://www.readbyqxmd.com/read/30005664/pxst-a-novel-vector-for-ta-cloning-and-blunt-end-cloning
#13
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
https://www.readbyqxmd.com/read/30005661/construction-of-a-trifunctional-cellulase-and-expression-in-saccharomyces-cerevisiae-using-a-fusion-protein
#14
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
https://www.readbyqxmd.com/read/29945600/membrane-bound-and-soluble-forms-of-an-nmda-receptor-extracellular-domain-retain-epitopes-targeted-in-auto-immune-encephalitis
#15
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
https://www.readbyqxmd.com/read/29945583/engineering-geobacillus-thermodenitrificans-to-introduce-cellulolytic-activity-expression-of-native-and-heterologous-cellulase-genes
#16
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
https://www.readbyqxmd.com/read/29945581/development-and-validation-of-an-integrated-dna-walking-strategy-to-detect-gmo-expressing-cry-genes
#17
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
https://www.readbyqxmd.com/read/29890980/tetanus-toxin-fragments-and-bcl-2-fusion-proteins-cytoprotection-and-retrograde-axonal-migration
#18
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
https://www.readbyqxmd.com/read/29859085/dna-assembly-with-error-correction-on-a-droplet-digital-microfluidics-platform
#19
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
https://www.readbyqxmd.com/read/29859082/heterologous-expression-and-biochemical-characterization-of-a-ghf9-endoglucanase-from-the-termite-reticulitermes-speratus-in-pichia-pastoris
#20
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
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