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A Feedback Mechanism to Control Apoptosis Occurs in the Digestive Gland of the Oyster Crassostrea gigas Exposed to the Paralytic Shellfish Toxins Producer Alexandrium catenella ArchiMer
Rolland, Jean-luc; Medhioub, Walid; Vergnes, Agnes; Abi-khalil, Celina; Savar, Veronique; Abadie, Eric; Masseret, Estelle; Amzil, Zouher; Laabir, Mohamed.
To better understand the effect of Paralytic Shellfish Toxins (PSTs) accumulation in the digestive gland of the Pacific oyster, Crassostrea gigas, we experimentally exposed individual oysters for 48 h to a PSTs producer, the dinoflagellate Alexandrium catenella. In comparison to the effect of the non-toxic Alexandrium tamarense, on the eight apoptotic related genes tested, Bax and BI.1 were significantly upregulated in oysters exposed 48 h to A. catenella. Among the five detoxification related genes tested, the expression of cytochrome P450 (CYP1A) was shown to be correlated with toxin concentration in the digestive gland of oysters exposed to the toxic dinoflagellate. Beside this, we observed a significant increase in ROS production, a decrease in...
Tipo: Text Palavras-chave: Shellfish; Toxin; Biomarker; Expression; Phytoplankton.
Ano: 2014 URL: http://archimer.ifremer.fr/doc/00225/33606/32003.pdf
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A novel expression vector for the secretion of abaecin in Bacillus subtilis BJM
Li,Li; Mu,Lan; Wang,Xiaojuan; Yu,Jingfeng; Hu,Ruiping; Li,Zhen.
ABSTRACT This study aimed to describe a Bacillus subtilis expression system based on genetically modified B. subtilis. Abaecin, an antimicrobial peptide obtained from Apis mellifera, can enhance the effect of pore-forming peptides from other species on the inhibition of bacterial growth. For the exogenous expression, the abaecin gene was fused with a tobacco etch virus protease cleavage site, a promoter Pglv, and a mature beta-glucanase signal peptide. Also, a B. subtilis expression system was constructed. The recombinant abaecin gene was expressed and purified as a recombinant protein in the culture supernatant. The purified abaecin did not inhibit the growth of Escherichia coli strain K88. Cecropin A and hymenoptaecin exhibited potent bactericidal...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Abaecin; Antimicrobial peptides; Bacillus subtilis; Expression.
Ano: 2017 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822017000400809
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Ammopiptanthus mongolicus stress-responsive NAC gene enhances the tolerance of transgenic Arabidopsis thaliana to drought and cold stresses Genet. Mol. Biol.
Pang,Xinyue; Xue,Min; Ren,Meiyan; Nan,Dina; Wu,Yaqi; Guo,Huiqin.
Abstract Drought and cold are the primary factors limiting plant growth worldwide. The Ammopiptanthus mongolicus NAC11 (AmNAC11) gene encodes a stress-responsive transcription factor. Expression of the AmNAC11 gene was induced by drought, cold and high salinity. The AmNAC11 protein was localized in the nucleus and plays an important role in tolerance to drought, cold and salt stresses. We also found that differential expression of AmNAC11 was induced in the early stages of seed germination and was related to root growth. When the AmNAC11 gene was introduced into Arabidopsis thaliana by an Agrobacterium-mediated method, the transgenic lines expressing AmNAC11 displayed significantly enhanced tolerance to drought and freezing stresses compared to wild-type...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Ammopiptanthus mongolicus; Expression; NAC; Tolerance; Transgene.
Ano: 2019 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572019000400624
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Application of n-dodecane as an oxygen vector to enhance the activity of fumarase in recombinant Escherichia coli: role of intracellular microenvironment BJM
Zhang,Sen; Song,Ping; Li,Shuang.
Abstract The effect of the intracellular microenvironment in the presence of an oxygen vector during expression of a fusion protein in Escherichia coli was studied. Three organic solutions at different concentration were chosen as oxygen vectors for fumarase expression. The addition of n-dodecane did not induce a significant change in the expression of fumarase, while the activity of fumarase increased significantly to 124% at 2.5% n-dodecane added after 9 h induction. The concentration of ATP increased sharply during the first 6 h of induction, to a value 7600% higher than that in the absence of an oxygen-vector. NAD/NADH and NADP/NADPH ratios were positively correlated with fumarase activity. n-Dodecane can be used to increase the concentration of ATP...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Oxygen vector; Energy; Fusion protein; Expression; Redox metabolism.
Ano: 2018 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822018000300662
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Cloning and expression analysis of rubredoxin from cold-treated banana leaves Phyton
Feng¹,RJ; Lu,LF; Yuan,KH; Cheng,P; Zhang,LL; Qi,JF; Ren,Y; Xu,XL; Zhang,XB; Zhou,LY; Zhang,YD.
A banana (Musa AAA, Cavendish subgroup cv. Brazil) cDNA encoding a putative rubredoxin-like protein (MaRd1) was obtained from total RNA isolated from cold-treated banana leaves using rapid amplification of cDNA ends (RACE) technique. MaRd1 cDNA contained 597 nucleotides encoding 198 amino acids in the open reading frame. MaRd1 protein showed 56% amino acid identity with that of Pyrococcus furiosus rubredoxin (P24297). A chloroplast transit peptide and a transmembrane region were detected at the N-terminus and the C-terminus, respectively, of the deduced amino acid sequence of MaRd1 gene. Southern blotting revealed the occurrence of at least two copies of MaRd1 in the banana genome. Real time quantitative RT-PCR analysis revealed that the expression of...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Rubredoxin; Expression; Banana; Stress response.
Ano: 2010 URL: http://www.scielo.org.ar/scielo.php?script=sci_arttext&pid=S1851-56572010000200008
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Cloning, bioinformatics analysis, and expression of the dust mite allergen Der f 5 of Dermatophagoides farinae BJMBR
Cui,Yubao; Zhou,Ying; Ma,Guifang; Yang,Li; Wang,Yungang; Shi,Weihong.
Crude extracts of house dust mites are used clinically for diagnosis and immunotherapy of allergic diseases, including bronchial asthma, perennial rhinitis, and atopic dermatitis. However, crude extracts are complexes with non-allergenic antigens and lack effective concentrations of important allergens, resulting in several side effects. Dermatophagoides farinae (Hughes; Acari: Pyroglyphidae) is one of the predominant sources of dust mite allergens, which has more than 30 groups of allergen. The cDNA coding for the group 5 allergen of D. farinae from China was cloned, sequenced and expressed. According to alignment using the VECTOR NTI 9.0 software, there were eight mismatched nucleotides in five cDNA clones resulting in seven incompatible amino acid...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Dermatophagoides farinae (Hughes); Der f 5; Cloning; Expression; Bioinformatics.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2012000800009
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Cloning, characterization and expression of a novel laccase gene Pclac2 from Phytophthora capsici BJM
Feng,Bao Zhen; Li,Peiqian.
Laccases are blue copper oxidases (E.C. 1.10.3.2) that catalyze the one-electron oxidation of phenolics, aromatic amines, and other electron-rich substrates with the concomitant reduction of O2 to H2O. A novel laccase gene pclac2 and its corresponding full-length cDNA were cloned and characterized from Phytophthora capsici for the first time. The 1683 bp full-length cDNA of pclac2 encoded a mature laccase protein containing 560 amino acids preceded by a signal peptide of 23 amino acids. The deduced protein sequence of PCLAC2 showed high similarity with other known fungal laccases and contained four copper-binding conserved domains of typical laccase protein. In order to achieve a high level secretion and full activity expression of PCLAC2, expression...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Phytophthora capsici; Laccase; Expression; Purification; Activity.
Ano: 2014 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000100050
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Cloning, sequence analysis, and expression of cDNA coding for the major house dust mite allergen, Der f 1, in Escherichia coli BJMBR
Cui,Y.; Zhou,P.; Peng,J.; Peng,M.; Zhou,Y.; Lin,Y.; Liu,L..
Our objective was to clone, express and characterize adult Dermatophagoides farinae group 1 (Der f 1) allergens to further produce recombinant allergens for future clinical applications in order to eliminate side reactions from crude extracts of mites. Based on GenBank data, we designed primers and amplified the cDNA fragment coding for Der f 1 by nested-PCR. After purification and recovery, the cDNA fragment was cloned into the pMD19-T vector. The fragment was then sequenced, subcloned into the plasmid pET28a(+), expressed in Escherichia coli BL21 and identified by Western blotting. The cDNA coding for Der f 1 was cloned, sequenced and expressed successfully. Sequence analysis showed the presence of an open reading frame containing 966 bp that encodes a...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Dermatophagoides farinae; Der f 1; Recombinant allergens; Cloning; Expression; Bioinformatics.
Ano: 2008 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2008000500006
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Cultivation of Pichia pastoris carrying the scFv anti LDL (-) antibody fragment. Effect of preculture carbon source BJM
Arias,Cesar Andres Diaz; Marques,Daniela de Araujo Viana; Malpiedi,Luciana Pellegrini; Maranhão,Andrea Queiroz; Parra,Dulcineia Abdalla Saes; Converti,Attilio; Pessoa Junior,Adalberto.
Abstract Antibodies and antibody fragments are nowadays among the most important biotechnological products, and Pichia pastoris is one of the most important vectors to produce them as well as other recombinant proteins. The conditions to effectively cultivate a P. pastoris strain previously genetically modified to produce the single-chain variable fragment anti low density lipoprotein (-) under the control of the alcohol oxidase promoter have been investigated in this study. In particular, it was evaluated if, and eventually how, the carbon source (glucose or glycerol) used in the preculture preceding cryopreservation in 20% glycerol influences both cell and antibody fragment productions either in flasks or in bioreactor. Although in flasks the volumetric...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Pichia pastoris; ScFv antibody fragment; Cryopreservation; Expression; Carbon source.
Ano: 2017 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822017000300419
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Design and expression of a retro doublet of cecropin with enhanced activity Electron. J. Biotechnol.
Díaz,Mauricio; Arenas,Gloria; Marshall,Sergio H.
Novel doublet molecules of cecropin A from Drosophila melanogaster were designed and constructed combining the regular (CECdir) with the inverted (CECret) coding sequence of the standard CEC A1 gene resulting in the following configurations: CECdir-CECret and CECret-CECdir. These two recombinant molecules were generated using a three-primer driven PCR reaction yielding composite single functional aminoacidic molecules with the coding sequences of CECdir linked in frame with the coding sequence of CECret and vice versa. In order to obtain these constructions, a retropeptide DNA-coding sequence was chemically synthesized to match the expected polarity of the newly generated CECret sequence. Both doublet antimicrobial peptides (drAMPs) were cloned in the T7...
Tipo: Journal article Palavras-chave: Antimicrobial peptides; Escherichia coli; Expression.
Ano: 2008 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000200006
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Differential expression of Zymomonas mobilis sucrase genes (sacB and sacC) in Escherichia coli and sucrase mutants of Zymomonas mobilis BABT
Gurunathan,Sangiliyandi; Gunasekaran,Paramasamy.
The sacB and sacC genes encoding levansucrase and extracellular sucrase respectively were independently subcloned in pBluescript (high copy number) and in Z. mobilis-E. coli shuttle vector, pZA22 (low copy number). The expression of these genes were compared under identical background of E. coli and Z. mobilis host. The level of sacB gene expression in E. coli was almost ten fold less than the expression of sacC gene, irrespective of the growth medium or the host strain. In Z. mobilis the expression of sacB and sacC genes was shown to be subject to carbon source dependent regulation. The transcript of sacB and sacC was three fold higher in cells grown on sucrose than in cells grown on glucose/fructose. Northern blot analysis revealed that the transcript...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Zymomonas mobilis; Escherichia coli; Expression; Levansucrase; Extracellular sucrase.
Ano: 2004 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132004000300001
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Effect of Photoperiod on the Egg Production, Plasma Luteinizing Hormone, Follicle-Stimulating Hormone, Gonadal Hormones, and Mrna Levels of Lh and Fsh in the Hypothalamic-Pituitary-Gonadal Axis of Pigeons Rev. Bras. Ciênc. Avic.
Wang,Y; Li,YB; Yang,HM; Wang,ZY.
ABSTRACT We investigated the egg production, changes in luteinizing hormone (LH), follicle-stimulating hormone (FSH), gonadal hormones, and their mRNA levels in the hypothalamic-pituitary-gonadal axis of White King pigeons submitted to different photoperiods. The treatments consisted of three photoperiods (8 h light (L):16 h dark (D), 12L:12D, and 16L:8D), with three replicates of twelve pairs of adult pigeons. The birds were exposed the photoperiods for 45 days. Egg production performance was recorded daily. Six pigeon pairs per replicate were selected for plasma collection, and six pigeon pairs per replicate for the resection of the hypothalamic-pituitary-gonadal (HPG) axis. Egg production was significantly improved by long-day lighting (16L:8D), while...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Day lighting; Expression; Hormone; Pigeon; Reproduction.
Ano: 2019 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-635X2019000400314
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Efficient expression and characterization of a cold-active endo-1, 4-β-glucanase from Citrobacter farmeri by co-expression of Myxococcus xanthus protein S Electron. J. Biotechnol.
Bai,Xi; Yuan,Xianjun; Wen,Aiyou; Li,Junfeng; Bai,Yunfeng; Shao,Tao.
Background: Cold-active endo-1, 4-β-glucanase (EglC) can decrease energy costs and prevent product denaturation in biotechnological processes. However, the nature EglC from C. farmeri A1 showed very low activity (800 U/L). In an attempt to increase its expression level, C. farmeri EglC was expressed in Escherichia coli as an N-terminal fusion to protein S (ProS) from Myxococcus xanthus. Results: A novel expression vector, pET(ProS-EglC), was successfully constructed for the expression of C. farmeri EglC in E. coli. SDS-PAGE showed that the recombinant protein (ProS-EglC) was approximately 60 kDa. The activity of ProS-EglC was 12,400 U/L, which was considerably higher than that of the nature EglC (800 U/L). ProS-EglC was active at pH 6.5-pH 8.0,...
Tipo: Journal article Palavras-chave: Cellulose degradation; Cellulose; Cold-active enzyme; Endoglucanases; Enzymatic properties; Escherichia coli; Expression; Novel expression vector; N-terminal fusion; Protein S-tag; Recombinant protein.
Ano: 2016 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000600012
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Etude des interactions hôte/parasite chez l'huître plate Ostrea edulis et son parasite Bonamia ostreae ArchiMer
Morga, Benjamin.
The history of the French oyster production highlights the fragility of this production against overexploitation and disease outbreaks. In particular, the production of flot oyster, Ostrea edulis, has decreased following the emergence of two parasitic diseases including bonamiosis. The means to fight against bonamiosis are relatively limited. They are mainly based on oyster health surveillance to limit the spread of the disease. However, the use of predictive models of disease progression in infected area would help to improve stock management and minimize the impact pathogens. Moreover the development of resistant animais could help to revive this production. These different approaches require appropriate diagnostic tools, a good knowledge of the life...
Tipo: Text Palavras-chave: Huître plate; Ostrea edulis; Bonamiose; Bonamia ostreae; Interaction hôte-parasite; Expression; Résistance; Hémocytes; Hybridation soustractive suppressive; Flat oyster; Ostrea edulis; Bonamiosis; Bonamia ostreae; Interaction host-parasite; Expression; Resistance; Haemocytes; Suppressive subtractive hybridization.
Ano: 2010 URL: http://archimer.ifremer.fr/doc/00055/16664/14157.pdf
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Expression analysis of the Arabidopsis thaliana AtSpen2 gene, and its relationship with other plant genes encoding Spen proteins Genet. Mol. Biol.
Solís-Guzmán,María Gloria; Argüello-Astorga,Gerardo; López-Bucio,José; Ruiz-Herrera,León Francisco; López-Meza,Joel; Sánchez-Calderón,Lenin; Carreón-Abud,Yazmín; Martínez-Trujillo,Miguel.
Abstract Proteins of the Split ends (Spen) family are characterized by an N-terminal domain, with one or more RNA recognition motifs and a SPOC domain. In Arabidopsis thaliana, the Spen protein FPA is involved in the control of flowering time as a component of an autonomous pathway independent of photoperiod. The A. thaliana genome encodes another gene for a putative Spen protein at the locus At4g12640, herein named AtSpen2. Bioinformatics analysis of the AtSPEN2 SPOC domain revealed low sequence similarity with the FPA SPOC domain, which was markedly lower than that found in other Spen proteins from unrelated plant species. To provide experimental information about the function of AtSpen2, A. thaliana plants were transformed with gene constructs of its...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Spen; Arabidopsis; Vascular bundle; Expression.
Ano: 2017 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572017000400643
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Expression and Localization of Olfactory Receptor AcerOr1 from Chinese Honey Bee, Apis cerana cerana (Hymenoptera, Apidae) BABT
Guo,Lina.
ABSTRACT Olfactory receptors are essential for recognition and detection of odor in honeybees. Although we have cloned and characterized the sequence of olfactory receptor AcerOr1 before, the tissue distribution and location of this gene in the nurse and the forager worker Apis cerana cerana were not very clear. To further investigate this information of AcerOr1 gene, we analyzed its expression and localization. The results showed that AcerOr1 mRNA was predominantly expressed in the antennae of nurse and forager bees, while the AcerOr1 protein was predominant in thorax, and its expression in antennae was higher in forager than in nurse. IHC revealed that AcerOr1 mainly localized in the olfactory neurons of the antennae. In addition, the staining intensity...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Apis cerana cerana; Olfactory receptor; AcerOr1; Expression; Localization; In vivo.
Ano: 2018 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132018000100440
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Expression of Babesia gibsoni thrombospondin-related adhesive protein in Toxoplasma gondii and evaluation of its antigenicity and immunogenicity OAK
Kumagai, A.; Zhang, G.; Jia, H.; Nakamura, C.; Zhang, H.; Goo, Y-K.; Aboge, G. A.; Terkawi, M. A.; Zhou, J.; Nishikawa, Y.; Xuan, X..
A gene encoding Babesia gibsoni thrombospondin-related adhesive protein (BgTRAP), known as a vaccine candidate, was stably expressed in Toxoplasma gondii (Tg/BgTRAP). The molecular weight and the antigenic reaction of recombinant BgTRAP expressed by the Tg/BgTRAP were similar to the original ones expressed by B. gibsoni. To evaluate the antigenicity of the recombinant BgTRAP expressed by T. gondii, the lysates of the recombinant parasite tachyzoites were intraperitoneally injected into mice. The serum collected from Tg/BgTRAP-immunized mouse could react to B. gibsoni parasites, while the serum collected from wild-type T. gondii tachyzoites (Tg/wt)-immunized mice did not. These results indicate that T. gondii could provide a new tool to produce foreign...
Palavras-chave: Toxoplasma gondii; Babesia gibsoni; BgTRAP; Expression; Vaccine.
Ano: 2016 URL: http://ir.obihiro.ac.jp/dspace/handle/10322/4496
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Expression of mouse beta defensin 2 in Escherichia coli and its broad-spectrum antimicrobial activity BJM
Gong,Tianxiang; Li,Wanyi; Wang,Yueling; Jiang,Yan; Zhang,Qiang; Feng,Wei; Jiang,Zhonghua; Li,Mingyuan.
Mature mouse beta defensin 2 (mBD2) is a small cationic peptide with antimicrobial activity. Here we established a prokaryotic expression vector containing the cDNA of mature mBD2 fused with thioredoxin (TrxA), pET32a-mBD2. The vector was transformed into Escherichia Coli (E. coli) Rosseta-gami (2) for expression fusion protein. Under the optimization of fermentation parameters: induce with 0.6 mM isopropylthiogalactoside (IPTG) at 34ºC in 2×YT medium and harvest at 6 h postinduction, fusion protein TrxA-mBD2 was high expressed in the soluble fraction (>95%). After cleaved fusion protein by enterokinase, soluble mature mBD2 was achieved 6 mg/L with a volumetric productivity. Purified recombinant mBD2 demonstrated clear broad-spectrum antimicrobial...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Mouse beta defensin 2; Expression; Purification; Antimicrobial activity.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822011000300043
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Expression, production, and renaturation of a functional single-chain variable antibody fragment (scFv) against human ICAM-1 BJMBR
Sun,H.; Wu,G.M.; Chen,Y.Y.; Tian,Y.; Yue,Y.H.; Zhang,G.L..
Intercellular adhesion molecule-1 (ICAM-1) is an important factor in the progression of inflammatory responses in vivo. To develop a new anti-inflammatory drug to block the biological activity of ICAM-1, we produced a monoclonal antibody (Ka=4.19×10−8 M) against human ICAM-1. The anti-ICAM-1 single-chain variable antibody fragment (scFv) was expressed at a high level as inclusion bodies in Escherichia coli. We refolded the scFv (Ka=2.35×10−7 M) by ion-exchange chromatography, dialysis, and dilution. The results showed that column chromatography refolding by high-performance Q Sepharose had remarkable advantages over conventional dilution and dialysis methods. Furthermore, the anti-ICAM-1 scFv yield of about 60 mg/L was higher with this method. The purity...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Intercellular adhesion molecule-1; Single-chain variable antibody fragment; Expression; Purification; Renaturation; Biological activity.
Ano: 2014 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2014000700540
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Gene expression pattern of digestive and antioxidant enzymes during the larval development of reared Atlantic bluefin tuna (ABFT), Thunnus thynnus L. ArchiMer
Mazurais, David; Coves, Denis; Papandroulakis, Nikos; Ortega, Aurelio; Desbruyeres, Elisabeth; Huelvan, Christine; Le Gall, Marie-madeleine; De La Gandara, Fernando; Cahu, Chantal.
The aim of this study was to determine whether mortality observed during the larval development of reared bluefin tuna (Thunnus thynnus) could be related to improper expression profiles of key genes involved in digestive or antioxidant response capabilities. Tuna larvae were sampled at hatching, 2, 5, 10, 15 and 20 dph (days post hatching) for the relative quantification of transcripts encoded by genes involved in digestive [trypsinogen 1 (TRYP1), alpha-amylase (AMY), aminopeptidase N (ANPEP)] and antioxidant [catalase (CAT)] functions. The levels of expression of ANPEP related to the development and maturation of intestinal function increased from 5 to 20 dph. Furthermore, AMY and TRYP1 genes, which are pancreatic enzymes implicated in carbohydrate and...
Tipo: Text Palavras-chave: Tuna; Gene; Expression; Development; Digestion; Antioxidant.
Ano: 2015 URL: https://archimer.ifremer.fr/doc/00171/28264/26521.pdf
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