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Recombinant expression

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https://www.readbyqxmd.com/read/28441623/identification-of-a-macrobrachium-nipponense-c-type-lectin-with-a-close-evolutionary-relationship-to-vertebrate-lectins
#1
Xin Huang, Tingting Li, Min Jin, Shaowu Yin, Wen Wang, Qian Ren
C-type lectins (CTLs) are involved in the innate immune defense of vertebrates and invertebrates against invading pathogens. This study cloned and characterized a novel C-type lectin (MnCTL) of the oriental river prawn, Macrobrachium nipponense. The cloned MnCTL cDNA encompasses an open reading frame of 774 nucleotides and encodes polypeptides of 257 residues. The deduced MnCTL protein contains a single carbohydrate recognition domain (CRD) with an EPN (Glu-Pro-Asn) motif in calcium-binding site 2. Phylogenetic analysis indicated that MnCTL has a closer evolutionary relationship with vertebrate lectins than with invertebrate lectins...
April 22, 2017: Molecular Immunology
https://www.readbyqxmd.com/read/28440881/overexpression-of-a-type-i-isopentenyl-pyrophosphate-isomerase-of-artemisia-annua-in-the-cytosol-leads-to-high-arteannuin-b-production-and-artemisinin-increase
#2
Dongming Ma, Gui Li, Fatima Alejos-Gonzalez, Yue Zhu, Zhen Xue, Aimin Wang, Hui Zhang, Xing Li, Hechun Ye, Hong Wang, Benye Liu, De-Yu Xie
We recently characterized a gene-terpene network that is associated with artemisinin biosynthesis in self-pollinated (SP) Artemisia annua, an effective antimalarial plant. We hypothesize that an alteration of gene expression in the network may improve the production of artemisinin and its precursors. In this study, we cloned an isopentenyl pyrophosphate isomerase (IPPI) cDNA, AaIPPI1, from Artemisia annua (Aa). The full-length cDNA encodes a type I IPPI containing a plastid transit peptide (PTP) at its amino terminus...
April 25, 2017: Plant Journal: for Cell and Molecular Biology
https://www.readbyqxmd.com/read/28440496/pu-1-serves-a-critical-role-in-the-innate-defense-against-aspergillus%C3%A2-fumigatus-via-dendritic-cell-associated-c-type-lectin-receptor-1-and-toll-like-receptors-2-and-4-in-thp-1-derived-macrophages
#3
Chenyang Liu, Min Wang, Wenkui Sun, Feng Cai, Shen Geng, Xin Su, Yi Shi
Aspergillus fumigatus (A. fumigatus) is one of the most common fungal pathogens of invasive pulmonary aspergillosis (IPA), which may be life threatening in immunocompromised individuals. The dendritic cell-associated C-type lectin receptor (Dectin-1), toll‑like receptor (TLR)-2 and TLR-4 are major pattern recognition receptors in alveolar macrophages that recognize A. fumigatus components. The PU.1 transcription factor is known to be important for the transcriptional control of these three receptors in mature macrophages...
April 24, 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28440490/microrna-15a-inhibition-protects-against-hypoxia-reoxygenation-induced-apoptosis-of-cardiomyocytes-by-targeting-mothers-against-decapentaplegic-homolog-7
#4
Yang Yang, Shiao Ding, Gaojun Xu, Fei Chen, Fangbao Ding
Myocardial ischemia/reperfusion (I/R) injury is a major pathological process in coronary heart disease and cardiac surgery, and is associated with aberrant microRNA (miR) expression. Previous studies have demonstrated that inhibition of miR-15a expression may ameliorate I/R‑induced myocardial injury. In the present study, the potential role and underlying mechanism of miR‑15a in hypoxia/reoxygenation‑induced apoptosis of cardiomyocytes was investigated. Myocardial I/R was simulated in cultured H9c2 cells by 24 h hypoxia followed by 24 h reoxygenation...
April 12, 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28440453/scrg1-suppresses-lps-induced-ccl22-production-through-erk1-2-activation-in-mouse-macrophage-raw264-7-cells
#5
Manabu Inoue, Junko Yamada, Emiko Aomatsu-Kikuchi, Kazuro Satoh, Hisatomo Kondo, Akira Ishisaki, Naoyuki Chosa
Recently, we identified the scrapie responsive gene 1 (SCRG1) secreted from mesenchymal stem cells (MSCs) and its receptor bone marrow stromal cell antigen 1 (BST1) as positive regulators of stem cell qualities such as self‑renewal, migration abilities, and osteogenic differentiation potential. Here, we examined the effect of the paracrine activity of SCRG1 in macrophages. The mouse macrophage‑like cell line Raw264.7 expressed BST1/β1 or BST1/β2 integrin as possible SCRG1 receptors. Unexpectedly, recombinant SCRG1 did not enhance cell proliferation, migration, or adhesion in these macrophages...
April 20, 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28440335/mycobacterium-tuberculosis-pe_pgrs41-enhances-the-intracellular-survival-of-m-smegmatis-within-macrophages-via-blocking-innate-immunity-and-inhibition-of-host-defense
#6
Wanyan Deng, Quanxin Long, Jie Zeng, Ping Li, Wenmin Yang, Xinchun Chen, Jianping Xie
The success of Mycobacterium tuberculosis (M. tuberculosis) as a pathogen is largely contributes to its ability to manipulate the host immune responses. The genome of M. tuberculosis encodes multiple immune-modulatory proteins, including several members of the multi-genic PE_PPE family. Despite of intense research, the roles of PE_PGRS proteins in mycobacterial pathogenesis remain elusive. The function of M. tuberculosis PE_PGRS41, characterized by an extended and unique C-terminal domain, was studied. Expression of PE_PGRS41 in Mycobacterium smegmatis, a non-pathogenic species intrinsically deficient of PE_PGRS, severely impaired the resistance of the recombinant to multiple stresses via altering the cell wall integrity...
April 25, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28439740/synthetic-cold-inducible-promoter-enhances-recombinant-protein-accumulation-during-agrobacterium-mediated-transient-expression-in-nicotiana-excelsior-at-chilling-temperatures
#7
I M Gerasymenko, Y V Sheludko
OBJECTIVES: To exploit cold-inducible biochemical processes beneficial for foreign mRNA transcription, translation and storage, as well as protein product stability, during Agrobacterium-mediated transient expression. RESULTS: The efficiency of three different 5'-regulatory sequences to achieve transient expression of the GFP-based reporter gene under chilling conditions (6-8 °C since the 3rd day post inoculation) was compared. We studied the upstream sequences of a cold-inducible Arabidopsis thaliana cor15a gene, the core element of 35S CaMV promoter fused to the TMV omega 5'-UTR, and the synthetic promoter including the 35S core sequence and two binding sites for cold-inducible CBF transcription factors (P_DRE::35S)...
April 24, 2017: Biotechnology Letters
https://www.readbyqxmd.com/read/28439515/endogenous-retroviruses-with-us-and-against-us
#8
REVIEW
Thomas J Meyer, Jimi L Rosenkrantz, Lucia Carbone, Shawn L Chavez
Mammalian genomes are scattered with thousands of copies of endogenous retroviruses (ERVs), mobile genetic elements that are relics of ancient retroviral infections. After inserting copies into the germ line of a host, most ERVs accumulate mutations that prevent the normal assembly of infectious viral particles, becoming trapped in host genomes and unable to leave to infect other cells. While most copies of ERVs are inactive, some are transcribed and encode the proteins needed to generate new insertions at novel loci...
2017: Frontiers in Chemistry
https://www.readbyqxmd.com/read/28439280/insights-on-structure-and-function-of-a-late-embryogenesis-abundant-protein-from-amaranthus-cruentus-an-intrinsically-disordered-protein-involved-in-protection-against-desiccation-oxidant-conditions-and-osmotic-stress
#9
Alma L Saucedo, Eric E Hernández-Domínguez, Luis A de Luna-Valdez, Angel A Guevara-García, Abraham Escobedo-Moratilla, Esaú Bojorquéz-Velázquez, Federico Del Río-Portilla, Daniel A Fernández-Velasco, Ana P Barba de la Rosa
Late embryogenesis abundant (LEA) proteins are part of a large protein family that protect other proteins from aggregation due to desiccation or osmotic stresses. Recently, the Amaranthus cruentus seed proteome was characterized by 2D-PAGE and one highly accumulated protein spot was identified as a LEA protein and was named AcLEA. In this work, AcLEA cDNA was cloned into an expression vector and the recombinant protein was purified and characterized. AcLEA encodes a 172 amino acid polypeptide with a predicted molecular mass of 18...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28439107/e-selectin-ligands-recognised-by-heca452-induce-drug-resistance-in-myeloma-which-is-overcome-by-the-e-selectin-antagonist-gmi-1271
#10
A Natoni, T A G Smith, N Keane, C McEllistrim, C Connolly, A Jha, M Andrulis, E Ellert, M S Raab, S V Glavey, L Kirkham-McCarthy, S K Kumar, S C Locatelli-Hoops, I Oliva, W E Fogler, J L Magnani, M E O'Dwyer
Multiple Myeloma (MM) is characterized by the clonal expansion and metastatic spread of malignant plasma cells to multiple sites in the bone marrow (BM). Recently, we implicated the sialyltransferase ST3Gal-6, an enzyme critical to the generation of E-selectin ligands, in MM BM homing and resistance to therapy. Since E-selectin is constitutively expressed in the BM microvasculature, we wished to establish the contribution of E-selectin ligands to MM biology. We report that functional E-selectin ligands are restricted to a minor subpopulation of MM cell lines which, upon expansion, demonstrate specific and robust interaction with recombinant E-selectin in vitro...
April 25, 2017: Leukemia: Official Journal of the Leukemia Society of America, Leukemia Research Fund, U.K
https://www.readbyqxmd.com/read/28438899/galectin-3-a-positive-regulator-of-leukocyte-recruitment-in-the-inflamed-microcirculation
#11
Beatrice R Gittens, Jennifer V Bodkin, Sussan Nourshargh, Mauro Perretti, Dianne Cooper
In vivo and ex vivo imaging were used to investigate the function of galectin-3 (Gal-3) during the process of leukocyte recruitment to the inflamed microcirculation. The cremasteric microcirculation of wild-type (C57BL/6), Gal-3(-/-), and CX3CR1(gfp/+) mice were assessed by intravital microscopy after PBS, IL-1β, TNF-α, or recombinant Gal-3 treatment. These cellular responses were investigated further using flow-chamber assays, confocal microscopy, flow cytometry, PCR analysis, and proteome array. We show that mechanisms mediating leukocyte slow rolling and emigration are impaired in Gal-3(-/-) mice, which could be because of impaired expression of cell adhesion molecules and an altered cell surface glycoproteome...
April 24, 2017: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/28438858/functional-roles-of-netrin-1-in-osteoblast-differentiation
#12
Tsuyoshi Sato, Shoichiro Kokabu, Yuichiro Enoki, Naoki Hayashi, Masahito Matsumoto, Mitsuhiko Nakahira, Masashi Sugasawa, Tetsuya Yoda
AIM: Recent studies have demonstrated that netrin-1 plays a vital role in bone metabolism. Previous studies have shown that osteoblasts produce netrin-1 which affects osteoclast differentiation. However, the role of netrin-1 in osteoblast differentiation is not well understood. In this study, we explored the roles of netrin-1 in osteoblasts. MATERIALS AND METHODS: Quantitative reverse-transcriptase polymerase chain reaction (qPCR), RNA interference for netrin receptors, the generation of netrin-1 plasmid, transfection of plasmids, and cell proliferation assay were performed...
May 2017: In Vivo
https://www.readbyqxmd.com/read/28438686/expression-purification-and-molecular-characterization-of-a-novel-endoglucanase-protein-from-bacillus-subtilis-sb13
#13
Xuefang Guan, Penglian Chen, Qingxian Xu, Lei Qian, Juqing Huang, Bin Lin
Bacillus subtilis strain SB13 which is isolated in our previous work was confirmed to produce endoglucanase. In this study, a novel endoglucanase gene (accession number: KX576676) was identified and cloned from SB13. Compared with other consensus sequence of reported endoglucanase genes in the GenBank database, this gene displays five differences (including T740C,A874G,A983G, T1210G and T1301C), which leading to five amino acid changes. Homology modeling has indicated that these five changes were located in the α-helix and random coil regions of the glycosyl hydrolase family 5 (GH5) domain, the random coil and β-sandwich of the type 3 carbohydrate-binding module (CBM3) domain, and the random coil domain...
April 21, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28438511/nitric-oxide-inhibits-the-mitochondrial-carnitine-acylcarnitine-carrier-through-reversible-s-nitrosylation-of-cysteine-136
#14
Annamaria Tonazzi, Nicola Giangregorio, Lara Console, Annalisa De Palma, Cesare Indiveri
S-nitrosylation of the mitochondrial Carnitine/Acylcarnitine transporter (CACT) has been investigated on the native and the recombinant proteins reconstituted in proteoliposomes, and on intact mitochondria. The widely-used NO-releasing compound, GSNO, strongly inhibited the antiport measured in proteoliposomes reconstituted with the native CACT from rat liver mitochondria or the recombinant rat CACT over-expressed in E. coli. Inhibition was reversed by the reducing agent dithioerithritol, indicating a reaction mechanism based on nitrosylation of Cys residues of the CACT...
April 21, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28438458/an-efficient-cumate-inducible-system-for-procyclic-and-bloodstream-form-trypanosoma-brucei
#15
Feng-Jun Li, Zhi-Shen Xu, Htay Mon Aye, Anaïs Brasseur, Zhao-Rong Lun, Kevin S W Tan, Cynthia Y He
In Trypanosoma brucei, the tetracycline-inducible system enables tightly-regulated, highly-efficient expression of recombinant proteins or double-stranded RNA in both procyclic and bloodstream form cells, providing useful molecular genetic tools to study gene functions. An alternative, vanillic acid-inducible system is recently described for procyclic T. brucei, providing ∼18-fold increase in GFP reporter expression upon induction (Sunter JD. Mol Biochem Parasitol. 2016, 207:45-48). Here we describe a cumate-inducible system that allows efficient, tunable gene expression showing >300-fold increase in GFP expression upon induction...
April 21, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28438456/pirab-protein-from-xenorhabdus-nematophila-hb310-exhibits-a-binary-toxin-with-insecticidal-activity-and-cytotoxicity-in-galleria-mellonella
#16
Qing Yang, Jie Zhang, Tianhui Li, Shen Liu, Ping Song, Ziyan Nangong, Qinying Wang
PirAB (Photorhabdus insect-related proteins, PirAB) toxin was initially found in the Photorhabdus luminescens TT01 strain and has been shown to be a binary toxin with high insecticidal activity. Based on GenBank data, this gene was also found in the Xenorhabdus nematophila genome sequence. The predicted amino acid sequence of pirA and pirB in the genome of X. nematophila showed 51% and 50% identity with those gene sequences from P. luminescens. The purpose of this experiment is to identify the relevant information for this toxin gene in X...
April 21, 2017: Journal of Invertebrate Pathology
https://www.readbyqxmd.com/read/28438410/vaccination-with-recombinant-modified-vaccinia-ankara-mva-viruses-expressing-single-african-horse-sickness-virus-vp2-antigens-induced-cross-reactive-virus-neutralising-antibodies-vnab-in-horses-when-administered-in-combination
#17
Nicola Mary Manning, Katarzyna Bachanek-Bankowska, Peter Paul Clement Mertens, Javier Castillo-Olivares
African horse sickness is a lethal viral disease of equids transmitted by biting midges of the Genus Culicoides. The disease is endemic to sub-Saharan Africa but outbreaks of high mortality and economic impact have occurred in the past in non-endemic regions of Africa, Asia and Southern Europe. Vaccination is critical for the control of this disease but only live attenuated vaccines are currently available. However, there are bio-safety concerns over the use of this type of vaccines, especially in non-endemic countries, and live attenuated vaccines do not have DIVA (Differentiation of Infected from Vaccinated Animals) capacity...
April 21, 2017: Vaccine
https://www.readbyqxmd.com/read/28438206/exogenous-gene-transfer-of-rab-3-8-small-gtpase-ameliorates-aberrant-lung-surfactant-homeostasis-in-ruby-rats
#18
Kazuhiro Osanai, Keisuke Nakase, Takashi Sakuma, Kazuaki Nishiki, Masafumi Nojiri, Ryo Kato, Masatoshi Saito, Yuki Fujimoto, Shiro Mizuno, Hirohisa Toga
BACKGROUND: Rab(3)8 small GTPase regulates intracellular transport in melanocytes and alveolar type II epithelial cells. Ruby rats carrying Rab(3)8 and other gene mutations exhibit oculocutaneous albinism, bleeding diathesis, and hence, are a rat model of human Hermansky-Pudlak syndrome (HPS). We previously showed that Long Evans Cinnamon (LEC) rats, one strain of the Ruby rats, developed aberrant lung surfactant homeostasis with remarkably enlarged lamellar bodies in alveolar type II cells...
April 24, 2017: Respiratory Research
https://www.readbyqxmd.com/read/28438205/expression-of-the-clonostachys-rosea-lactonohydrolase-gene-by-lactobacillus-reuteri-to-increase-its-zearalenone-removing-ability
#19
Wen-Chun Yang, Tsui-Chun Hsu, Kuan-Chen Cheng, Je-Ruei Liu
BACKGROUND: Mycotoxins are secondary metabolites produced by filamentous fungi that can contaminate agricultural crops in the field as well as during harvest, transportation, processing, or storage. Zearalenone (ZEN), a non-steroidal estrogenic mycotoxin, produced by Fusarium species, has been shown to be associated with reproductive disorders in farm animals and to a lesser extent in hyperoestrogenic syndromes in humans. Thus, the decontamination of ZEN in foods and feeds is an important issue...
April 24, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28438014/characterization-of-a-glutamate-decarboxylase-gad-from-enterococcus-avium-m5-isolated-from-jeotgal-a-fermented-korean-sea-food
#20
Kang Wook Lee, Jae Min Shim, Zhuang Yao, Jeong A Kim, Hyun-Jin Kim, Jeong Hwan Kim
To develop starters for the production of functional foods or materials, lactic acid bacteria producing γ-aminobutyric acid (GABA) were screened from jeotgals, Korean fermented sea foods. One isolate producing high amount of GABA from monosodium L-glutamate (MSG) was identified as Enterococcus avium by 16S rRNA gene sequencing. E. avium M5 produced 18.47 ± 1.26 mg/ml GABA when incubated for 48 h at 37°C in MRS broth with MSG (3%, w/v). A gadB gene encoding glutamate decarboxylase (GAD) was cloned and overexpressed in E...
April 25, 2017: Journal of Microbiology and Biotechnology
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