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https://www.readbyqxmd.com/read/29782978/a-potent-chitinase-from-bacillus-subtilis-for-the-efficient-bioconversion-of-chitin-containing-wastes
#1
Di Wang, Anjie Li, Hongyu Han, Tian Liu, Qing Yang
Bioconversion of chitin-containing waste to N-acetyl-d-glucosamine (GlcNAc) is of economic interest. Highly efficient chitinases are thus desirable. Bacillus subtilis is a well-known microorganism for industrial protein production and biocontrol, however its chitinolytic enzyme has not been studied in detail. In this study, BsChi from B. subtilis was recombinantly expressed in Escherichia coli and characterized as a potent chitinase for degrading crystalline chitin substrates such as α-chitin, β-chitin and crab shells...
May 18, 2018: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/29782916/identification-of-amino-acid-residues-in-infectious-hematopoietic-necrosis-virus-ihnv-nv-protein-necessary-for-viral-replication-and-pathogenicity
#2
Yang Wu, Li Wang, Tiantian Guo, Yanping Jiang, Xinyuan Qiao, Li Sun, Min Liu, Lijie Tang, Yigang Xu, Yijing Li
Our previous studies demonstrated that the nonstructural NV protein of infectious hematopoietic necrosis virus (IHNV) was essential for efficient viral replication and pathogenicity, and that the amino acid residues 32 EGDL35 of the NV protein were responsible for nuclear localization, and played important roles in suppressing IFN and inhibiting NF-κB activity. However, little is known about the influence of 32 EGDL35 on IHNV replication and pathogenicity. In the present study, two recombinant IHNV strains with deletions of NV 32 EGDL35 were generated and the effect on IHNV replication and pathogenicity was explored...
May 18, 2018: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/29782825/characterization-of-an-n-terminal-mutant-of-%C3%AE-a-crystallin-%C3%AE-a-r21q-associated-with-congenital-cataract
#3
Ashutosh S Phadte, Puttur Santhoshkumar, K Krishna Sharma
Several mutations associated with congenital cataracts in human beings target conserved arginine residues in αA-crystallin. The N-terminal region of αA-crystallin is a "mutational hotspot," with multiple cataract-related mutations reported in this region. Two mutations at arginine 21 in the N-terminal domain of αA-crystallin - αA-R21L and αA-R21W have been associated with congenital cataract. A third mutant of R21, αA-R21Q, was recently identified to be associated with congenital cataract in a South Australian family...
May 18, 2018: Experimental Eye Research
https://www.readbyqxmd.com/read/29782238/generation-of-a-lineage-ii-powassan-virus-deer-tick-virus-cdna-clone-assessment-of-flaviviral-genetic-determinants-of-tick-and-mosquito-vector-competence
#4
Joan L Kenney, Michael Anishchenko, Meghan Hermance, Hannah Romo, Ching-I Chen, Saravanan Thangamani, Aaron C Brault
The Flavivirus genus comprises a diverse group of viruses that utilize a wide range of vertebrate hosts and arthropod vectors. The genus includes viruses that are transmitted solely by mosquitoes or vertebrate hosts as well as viruses that alternate transmission between mosquitoes or ticks and vertebrates. Nevertheless, the viral genetic determinants that dictate these unique flaviviral host and vector specificities have been poorly characterized. In this report, a cDNA clone of a flavivirus that is transmitted between ticks and vertebrates (Powassan lineage II, deer tick virus [DTV]) was generated and chimeric viruses between the mosquito/vertebrate flavivirus, West Nile virus (WNV), were constructed...
May 21, 2018: Vector Borne and Zoonotic Diseases
https://www.readbyqxmd.com/read/29782165/linked-cluster-formulation-of-electron-hole-interaction-kernel-in-real-space-representation-without-using-unoccupied-states
#5
Michael G Bayne, Jeremy A Scher, Benjamin H Ellis, Arindam Chakraborty
Electron-hole or quasiparticle representation plays a central role in describing electronic excitations in many-electron systems. For charge-neutral excitation, the electron-hole interaction kernel is the quantity of interest for calculating important excitation properties such as optical gap, optical spectra, electron-hole recombination and electron-hole binding energies. The electron-hole interaction kernel can be formally derived from the density-density correlation function using both Green's function and TDDFT formalism...
May 21, 2018: Journal of Chemical Theory and Computation
https://www.readbyqxmd.com/read/29781997/antimicrobial-peptides-produced-by-selective-pressure-incorporation-of-non-canonical-amino-acids
#6
Jessica H Nickling, Tobias Baumann, Franz-Josef Schmitt, Maike Bartholomae, Oscar P Kuipers, Thomas Friedrich, Nediljko Budisa
Nature has a variety of possibilities to create new protein functions by modifying the sequence of the individual amino acid building blocks. However, all variations are based on the 20 canonical amino acids (cAAs). As a way to introduce additional physicochemical properties into polypeptides, the incorporation of non-canonical amino acids (ncAAs) is increasingly used in protein engineering. Due to their relatively short length, the modification of ribosomally synthesized and post-translationally modified peptides by ncAAs is particularly attractive...
May 4, 2018: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29781203/the-long-non-coding-rna-transcriptome-landscape-in-cho-cells-under-batch-and-fed-batch-conditions
#7
Davide Vito, C Mark Smales
The role of non-coding RNAs in determining growth, productivity and recombinant product quality attributes in Chinese hamster ovary (CHO) cells has received much attention in recent years, exemplified by studies into microRNAs in particular. However, other classes of non-coding RNAs have received less attention. One such class are the non-coding RNAs known collectively as long non-coding RNAs (lncRNAs). We have undertaken the first landscape analysis of the lncRNA transcriptome in CHO using a mouse based microarray that also provided for the surveillance of the coding transcriptome...
May 21, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29781190/in-situ-study-of-the-functions-of-bacterioruberin-in-the-dual-chromophore-photoreceptor-archaerhodopsin-4
#8
Chao Sun, Xiaoyan Ding, Haolin Cui, Yanan Yang, Sijin Chen, Anthony Watts, Xin Zhao
While certain archaeal ion pumps have been shown to contain two chromophores, retinal and carotenoid bacterioruberin, the functions of the bacterioruberin have not been well explored. To address this research gap, the recombinant archaerhodopsin-4 (aR4), either with retinal chromophore only or with both retinal and bacterioruberin chromophores, was successfully expressed together with endogenous lipids in H. salinarum L33 and MPK409 respectively. In situ solid-state NMR, supported by molecular spectroscopy and functional assays, revealed for the first time that the retinal thermal equilibrium in the dark-adapted state is modulated by bacterioruberin binding via a cluster of aromatic residues on helix E...
May 20, 2018: Angewandte Chemie
https://www.readbyqxmd.com/read/29781132/egr2-mediates-the-differentiation-of-mouse-uterine-stromal-cells-responsiveness-to-hb-egf-during-decidualization
#9
Liang Yue, Hai-Fan Yu, Zhan-Qing Yang, Xue-Chao Tian, Lian-Wen Zheng, Bin Guo
Although Egr2 is involved in regulating the folliculogenesis and ovulation, there is almost no data describing its physiological function in embryo implantation and decidualization. Here, we showed that Egr2 mRNA was distinctly accumulated in subluminal stromal cells around implanting blastocyst on day 5 of pregnancy as well as in estrogen-activated implantation uterus. Estrogen induced the expression of Egr2 in uterine epithelia. Elevated expression of Egr2 mRNA was also observed in the decidual cells. Silencing of Egr2 by specific siRNA weakened the proliferation of uterine stromal cells and reduced the expression of Ccnd1, Ccnd3, Cdk4, and Cdk6...
May 20, 2018: Journal of Experimental Zoology. Part B, Molecular and Developmental Evolution
https://www.readbyqxmd.com/read/29780379/point-mutation-of-a-non-elastase-binding-site-in-human-%C3%AE-1-antitrypsin-alters-its-anti-inflammatory-properties
#10
Yotam Lior, Mariana Zaretsky, David E Ochayon, Diana Lotysh, Boris M Baranovski, Ronen Schuster, Ofer Guttman, Amir Aharoni, Eli C Lewis
Introduction: Human α1-antitrypsin (hAAT) is a 394-amino acid long anti-inflammatory, neutrophil elastase inhibitor, which binds elastase via a sequence-specific molecular protrusion (reactive center loop, RCL; positions 357-366). hAAT formulations that lack protease inhibition were shown to maintain their anti-inflammatory activities, suggesting that some attributes of the molecule may reside in extra-RCL segments. Here, we compare the protease-inhibitory and anti-inflammatory profiles of an extra-RCL mutation (cys232pro) and two intra-RCL mutations (pro357cys, pro357ala), to naïve [wild-type (WT)] recombinant hAAT, in vitro , and in vivo ...
2018: Frontiers in Immunology
https://www.readbyqxmd.com/read/29780024/the-neutrophil-mobilizing-cytokine-interleukin-26-in-the-airways-of-long-term-tobacco-smokers
#11
Karlhans Fru Che, Ellen Tufvesson, Sara Tengvall, Elisa Lappi-Blanco, Riitta Kaarteenaho, Bettina Levänen, Marie Ekberg, Annelie Brauner, Åsa M Wheelock, Leif Bjermer, C Magnus Sköld, Anders Lindén
Long-term tobacco smokers with chronic obstructive pulmonary disease (COPD) or chronic bronchitis display an excessive accumulation of neutrophils in the airways; an inflammation that responds poorly to established therapy. Thus, there is a need to identify new molecular targets for the development of effective therapy. Here, we hypothesized that the neutrophil-mobilizing cytokine interleukin (IL)-26 (IL-26) is involved in airway inflammation amongst long-term tobacco smokers with or without COPD, chronic bronchitis or colonization by pathogenic bacteria...
May 23, 2018: Clinical Science (1979-)
https://www.readbyqxmd.com/read/29779329/-mechanisms-of-recombinant-adenovirus-mediated-sd-ha-fusion-protein-proliferation-inhibition-and-induced-apoptosis-of-k562-cells
#12
Y Huang, P Zhang, L Du, M Gui, W L Feng, Z Peng
Objective: To investigate whether fusion protein SD-HA could regulate its downstream signaling molecule activity by competing with the phospho-BCR-ABL Y177 site, and its mechanisms to inhibit proliferation and induce apoptosis of K562 cells. Methods: Co-immunoprecipitation interaction technology analysis of fusion protein SD-HA functioned by potently binding to the phospho-BCR-ABL Y177 site, Ras, MAPK and Akt activities were observed in the Ad5F35-SD-HA-treated cells. Western blot analyses of SD-HA fusion protein on cell membrane receptor pathway to death cascade caspase-8, caspase-3 and PRAP were performed...
April 14, 2018: Zhonghua Xue Ye Xue za Zhi, Zhonghua Xueyexue Zazhi
https://www.readbyqxmd.com/read/29779183/molecular-characterization-and-potential-synthetic-applications-of-gh1-%C3%AE-glucosidase-from-higher-termite-microcerotermes-annandalei
#13
Siriphan Arthornthurasuk, Wantha Jenkhetkan, Eukote Suwan, Daranee Chokchaichamnankit, Chantragan Srisomsap, Pakorn Wattana-Amorn, Jisnuson Svasti, Prachumporn T Kongsaeree
A novel β-glucosidase from higher termite Microcerotermes annandalei (MaBG) was obtained via a screening method targeting β-glucosidases with increased activities in the presence of glucose. The purified natural MaBG showed a subunit molecular weight of 55 kDa and existed in a native form as a dimer without any glycosylation. Gene-specific primers designed from its partial amino acid sequences were used to amplify the corresponding 1,419-bp coding sequence of MaBG which encodes a 472-amino acid glycoside hydrolase family 1 (GH1) β-glucosidase...
May 19, 2018: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/29779122/heterologous-expression-of-talaromyces-emersonii-cellobiohydrolase-cel7a-in-trichoderma-reesei-increases-the-efficiency-of-corncob-residues-saccharification
#14
Ningning Sun, Yuanchao Qian, Weiwei Wang, Yaohua Zhong, Meixue Dai
OBJECTIVE: Improve the hydrolysis efficiency of the Trichoderma reesei cellulase system by heterologously expressing cellobiohydrolase Cel7A (Te-Cel7A) from the thermophilic fungus Talaromyces emersonii. RESULTS: Te-Cel7A was expressed in T. reesei under control of the cdna1 promoter and the generated transformant QTC14 could successfully secrete Te-Cel7A into the supernatant using glucose as carbon source. The recombinant Te-Cel7A had a temperature optimum at 65 °C and an optimal pH of 5, which were similar to those from the native host...
May 19, 2018: Biotechnology Letters
https://www.readbyqxmd.com/read/29779115/the-first-recombinant-viper-three-finger-toxins-inhibition-of-muscle-and-neuronal-nicotinic-acetylcholine-receptors
#15
Ya V Makarova, E V Kryukova, I V Shelukhina, D S Lebedev, T V Andreeva, D Yu Ryazantsev, S V Balandin, T V Ovchinnikova, V I Tsetlin, Yu N Utkin
Genes encoding two three-finger toxins TFT-AF and TFT-VN, nucleotide sequences of which were earlier determined by cloning cDNA from venom glands of vipers Azemiops feae and Vipera nikolskii, respectively, were expressed for the first time in E. coli cells. The biological activity of these toxins was studied by electrophysiological techniques, calcium imaging, and radioligand analysis. It was shown for the first time that viper three-finger toxins are antagonists of nicotinic acetylcholine receptors of neuronal and muscle type...
March 2018: Doklady. Biochemistry and Biophysics
https://www.readbyqxmd.com/read/29778647/expression-of-recombinant-proteins-in-insect-and-mammalian-cells
#16
REVIEW
Edward A McKenzie, W Mark Abbott
Purified recombinant proteins are key reagents in academic and industrial research. The ability to make these proteins quickly often relies on the availability of higher eukaryotic cell hosts such as insect and mammalian cells where there is a very wide range of post-translational modifications, protein folding and trafficking pathways. This enables the generation of many proteins that cannot be made in microbial hosts. In this article we outline some of the most commonly used methods to express recombinant proteins in insect and mammalian cells...
May 17, 2018: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/29778203/evaluation-of-the-immune-response-afforded-by-a-subunit-vaccine-candidate-against-bluetongue-virus-in-mice-and-sheep
#17
Darien Kheder Ali Mohamed, Junzheng Du, Shandian Gao, Zhancheng Tian, Guorui Zhang, Dexuan Huang, Rongsheng Du, Biao Kang, Guangyuan Liu, Jianxun Luo, Hong Yin
Bluetongue virus (BTV), a vector-borne pathogen, is the causative agent of bluetongue disease in ruminants. In view of the recent emergence of BTV in regions previously known to be free from the disease and/or specific serotypes or strains, optimization of the currently available vaccination strategies to control the spread of vector-borne bluetongue is crucial. The main objective of the current study was to develop a subunit vaccine candidate targeting BTV-16, a strain previously isolated in China from sheep with obvious clinical signs...
June 2018: Veterinary Microbiology
https://www.readbyqxmd.com/read/29778186/identification-of-antigenic-epitopes-of-monoclonal-antibodies-against-the-vp2-protein-of-the-25-serotype-of-bluetongue-virus
#18
Shuangyu Xie, Yiming Shi, Ruyue Gong, Wen Cui, Yanping Jiang, Min Liu, Li Wang, Han Zhou, Xinyuan Qiao, Yijing Li, Yigang Xu, Lijie Tang
Serious outbreaks of bluetongue, an arbovirus of domestic and wild ruminants caused by bluetongue virus serotypes (BTV), have occurred around the world. More than 27 distinct serotypes are recognized throughout the world. A new virus, BTV-25 (Toggenburg orbivirus [TOV]), was first detected in Switzerland, and has not yet been found in China. VP2 is an important outer shell protein that defines BTV serotypes and is, therefore, an ideal target antigen for serotype identification. To produce a monoclonal antibody against VP2 of BTV-25, the segment 2 gene was divided into three segments, cloned into pET-28a (+) and pMAL-c5X vectors, and the protein was expressed in E...
June 2018: Veterinary Microbiology
https://www.readbyqxmd.com/read/29778179/recombinant-adenovirus-delivered-soluble-cd163-and-sialoadhesin-receptors-protected-pigs-from-porcine-reproductive-and-respiratory-syndrome-virus-infection
#19
Wenlong Xia, Zhi Wu, Changming Guo, Shanyuan Zhu, Xinyu Zhang, Xiaoli Xia, Huaichang Sun
Porcine reproductive and respiratory syndrome (PRRS) is one of the most important swine diseases affecting pig industry worldwide. Sialoadehesin (Sn) and CD163 are the two specific receptors for PRRSV infection of porcine alveolar macrophages. Our previous study showed that the soluble Sn receptor Sn4D-Fc and soluble CD163 receptor SRCR59-Fc expressed by the two recombinant adenoviral (rAd) vectors have an additive anti-PRRSV effect in vitro. In the present study, rAd-Sn4D-Fc and rAd-SRCR59-Fc were inoculated into pigs, and the efficient expression of Sn4D-Fc and SRCR59-Fc proteins was detected by ELISA...
June 2018: Veterinary Microbiology
https://www.readbyqxmd.com/read/29778095/establishing-an-innovative-carbohydrate-metabolic-pathway-for-efficient-production-of-2-keto-l-gulonic-acid-in-ketogulonicigenium-robustum-initiated-by-intronic-promoters
#20
Cai-Yun Wang, Ye Li, Zi-Wei Gao, Li-Cheng Liu, Meng-Yue Zhang, Tian-Yuan Zhang, Chun-Fu Wu, Yi-Xuan Zhang
BACKGROUND: 2-Keto-L-gulonic acid (2-KGA), the precursor of vitamin C, is currently produced by two-step fermentation. In the second step, L-sorbose is transformed into 2-KGA by the symbiosis system composed of Ketogulonicigenium vulgare and Bacillus megaterium. Due to the different nutrient requirements and the uncertain ratio of the two strains, the symbiosis system significantly limits strain improvement and fermentation optimization. RESULTS: In this study, Ketogulonicigenium robustum SPU_B003 was reported for its capability to grow well independently and to produce more 2-KGA than that of K...
May 19, 2018: Microbial Cell Factories
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