Refolding and characterization of a yeast dehydrodolichyl diphosphate synthase overexpressed in Escherichia coli.

abstract：:Enteroaggregative Escherichia coli (EAEC) heat-stable enterotoxin 1 (EAST1) is a 4.1k Da protein originally discovered in EAEC but known to be scattered in other diarrheagenic E. coli as well, possibly causing diarrhea in humans and animals. We report for the first time a method to express and purify EAST1 using the G...

journal_title：Protein expression and purification

authors： Ménard LP,Lussier JG,Lépine F,Paiva de Sousa C,Dubreuil JD

更新日期：2004-02-01 00:00:00

abstract：:This is the first report of an insect esterase efficiently expressed in the methylotrophic yeast Pichia pastoris (so far insect esterases have been produced only in the baculovirus system). Having isolated a Tribolium castaneum carboxylesterase cDNA (TCE), we were initially unable to express it in Escherichia coli or ...

journal_title：Protein expression and purification

authors： Delroisse JM,Dannau M,Gilsoul JJ,El Mejdoub T,Destain J,Portetelle D,Thonart P,Haubruge E,Vandenbol M

更新日期：2005-08-01 00:00:00

abstract：:Membrane proteins, including solute transporters play crucial roles in cellular function and have been implicated in a variety of important diseases, and as such are considered important targets for drug development. Currently the drug discovery process is heavily reliant on the structural and functional information d...

journal_title：Protein expression and purification

authors： Maggioni A,Hadley B,von Itzstein M,Tiralongo J

更新日期：2014-09-01 00:00:00

abstract：:A gene encoding a glutamate-specific endopeptidase (GSE) from Bacillus licheniformis (BL) has been cloned in Escherichia coli cells. The recombinant protein was expressed as cytoplasmic insoluble inclusion bodies. Immobilized metal affinity chromatography was employed to purify the protein, and then a 27-kDa GSE inter...

journal_title：Protein expression and purification

authors： Ye W,Liu J,Wang H,Wang J,Wang X

更新日期：2012-03-01 00:00:00

abstract：:Glucose-6-phosphate dehydrogenase was purified from human placenta using DEAE-Sepharose fast flow, 2',5'-ADP Sepharose 4B column chromatography, and chromatofocusing on PBE 94 with PB 74. The enzyme was purified with 62% yield and had a specific activity of 87 units per milligram protein. The pH optimum was determined...

journal_title：Protein expression and purification

authors： Aksoy Y,Ogüs IH,Oauzer N

更新日期：2001-03-01 00:00:00

abstract：:The gene encoding S. cerevisiae Kex2 protease derivative Kex2-667 (encoding the N-terminal 20th to 667th amino acid residues of Kex2 protease, containing the propeptide, catalytic domain, P domain and Ser/Thr enrichment region) and its 225th amino acid residue mutant K225L were overexpressed in Pichia pastoris. Protea...

journal_title：Protein expression and purification

authors： Yang F,Liu L,Liu Y,Li S

更新日期：2020-12-01 00:00:00

abstract：:The second enzyme of the naphthalene degradation pathway in Pseudomonas putida G7 is NahB, a dehydrogenase that converts cis-1,2-dihydroxy-1,2-dihydronaphthalene to 1,2-dihydroxynaphthalene. We report the cloning, optimization of expression, purification, kinetic studies and preliminary structural characterization of ...

journal_title：Protein expression and purification

authors： Costa DMA,Costa MAF,Guimarães SL,Coitinho JB,Gómez SV,Brandão TADS,Nagem RAP

更新日期：2017-04-01 00:00:00

abstract：:Autophagy is the process of degradation of intracellular proteins through the lysosome. Members of the tripartite motif (TRIM) proteins have shown to directly recognize autophagic cargo and also to act as a hub for the phagophore nucleation complex. The TRIM proteins are classically characterized by the presence of an...

journal_title：Protein expression and purification

authors： Guimarães DS,Gomes MD

更新日期：2018-03-01 00:00:00

abstract：:A full-length hexokinase cDNA was cloned from Solanum chacoense, a wild relative of the cultivated potato. Analysis of the predicted primary sequence suggested that the protein product, ScHK2, may be targeted to the secretory pathway and inserted in the plant plasma membrane, facing the cytosol. ScHK2 was expressed as...

journal_title：Protein expression and purification

authors： Claeyssen E,Wally O,Matton DP,Morse D,Rivoal J

更新日期：2006-05-01 00:00:00

abstract：:Amidase signature (AS) family amidases are known to exhibit broad substrate specificity. According to the available genome sequence data, a novel AS family amidase, Pl-Ami, was identified and cloned from the genome of Parvibaculum lavamentivorans ZJB14001. The recombinant amidase was overexpressed in Escherichia coli ...

journal_title：Protein expression and purification

authors： Wu ZM,Zheng RC,Zheng YG

更新日期：2017-01-01 00:00:00

abstract：:The mRNA encoding the 51-kDa subunit of 6-phosphogluconate dehydrogenase (6PGDH) from sheep liver was reverse-transcribed and amplified. The resulting cDNA was reamplified in N-terminal and C-terminal segments and spliced to generate a full-length clone, and an internal cDNA fragment was also amplified. The full-lengt...

journal_title：Protein expression and purification

authors： Chooback L,Price NE,Karsten WE,Nelson J,Sundstrom P,Cook PF

更新日期：1998-07-01 00:00:00

abstract：:Five variants of human β-defensins (HBDs) were expressed in Escherichia coli using two vector systems (pET28a(+) and pQE30) with inducible expression by IPTG. The last vector has not been previously reported as an expression system for HBDs. The recombinant peptides were different in their lengths and overall charge. ...

journal_title：Protein expression and purification

authors： Corrales-Garcia L,Ortiz E,Castañeda-Delgado J,Rivas-Santiago B,Corzo G

更新日期：2013-05-01 00:00:00

abstract：:Plants possess very large numbers of biosynthetic cytochrome P450 enzymes. In spite of the importance of these enzymes for the synthesis of bioactive plant secondary metabolites, only two plant P450 structures has been obtained to date. Isoflavone synthase (IFS) is a membrane-associated cytochrome P450 enzyme catalyzi...

journal_title：Protein expression and purification

authors： Chang Z,Wang X,Wei R,Liu Z,Shan H,Fan G,Hu H

更新日期：2010-12-04 00:00:00

abstract：:Mitochondrial 2,4-dienoyl-CoA reductase is a key enzyme for the beta-oxidation of unsaturated fatty acids. The cDNA of the full-length human mitochondrial 2,4-dienoyl-CoA reductase was previously cloned as pUC18::DECR. PCR methodologies were used to subclone the genes encoding various truncated human mitochondrial 2,4...

journal_title：Protein expression and purification

authors： Chu X,Yu W,Chen G,Li D

更新日期：2003-10-01 00:00:00

abstract：:Mitochondria-mediated apoptosis (programmed cell death) involves a sophisticated signaling and regulatory network that is regulated by the Bcl-2 protein family. Members of this family have either pro- or anti-apoptotic functions. An important pro-apoptotic member of this family is the cytosolic Bax. This protein is cr...

journal_title：Protein expression and purification

authors： Dingeldein APG,Lindberg MJ,Ådén J,Zhong X,Stoll R,Gröbner G

更新日期：2019-06-01 00:00:00

abstract：:A long-lasting recombinant human albumin-linker-erythropoietin (EPO) is a human albumin gene fused to the N-terminal of EPO with a (GGSGG)(n)-repeated linker inserted between albumin and EPO. Albumin-EPO fusion genes were co-transfected with the dhfr gene. Albumin-EPO fusion protein has three kinds of sub-types (IALE,...

journal_title：Protein expression and purification

authors： Joung CH,Shin JY,Koo JK,Lim JJ,Wang JS,Lee SJ,Tan HK,Kim SL,Lim SM

更新日期：2009-12-01 00:00:00

abstract：:Most parasitic protozoa lack the de novo purine biosynthetic pathway and rely exclusively on the salvage pathway for their purine nucleotide requirements. Enzymes of the salvage pathway are, therefore, candidate drug targets. We have cloned the Plasmodium falciparum adenylosuccinate synthetase gene. In the parasite, a...

journal_title：Protein expression and purification

authors： Jayalakshmi R,Sumathy K,Balaram H

更新日期：2002-06-01 00:00:00

abstract：:A soluble N-terminal domain of the Escherichia coli adenylyl transferase (ATase) is responsible for deadenylylation activity of the intact enzyme. Previous studies of the deadenylylation activity have involved a fragment, AT-N423 (residues 1 to 423), which was extended by 17 amino acids to give AT-N440. This new domai...

journal_title：Protein expression and purification

authors： Xu Y,Wen D,Clancy P,Carr PD,Ollis DL,Vasudevan SG

更新日期：2004-03-01 00:00:00

abstract：:Recombinant OSU VP4 protein, and outer capsid antigen of porcine rotavirus, was purified to a high level from the spent broth of baculovirus-infected Spodoptera frugiperda insect cells. Initial clarification of the broth with a 0-60% ammonium sulfate cut retained 93% of the total VP4. Q-sepharose ion exchange chromato...

journal_title：Protein expression and purification

authors： Juarbe-Osorio LG,Gorziglia M,Betenbaugh MJ

更新日期：1993-06-01 00:00:00

abstract：:Protein convertase 1/3 is a serine endoproteinase present in the regulated secretory pathway of endocrine and neuroendocrine cells. It is responsible for the processing of numerous prohormones and proneuropeptides into their biologically active moieties, often following cleavage at pairs of basic residues. The determi...

journal_title：Protein expression and purification

authors： Rabah N,Gauthier DJ,Gauthier D,Lazure C

更新日期：2004-10-01 00:00:00

abstract：:Fatty acid desaturases are enzymes that introduce double bonds into fatty acyl chains, among which stearoyl-acyl carrier protein desaturase (S-ACP-DES) was widely distributed in the plant kingdom. We cloned the cDNA coding for fab2/ssi2, an S-ACP-DES from Arabidopsis thaliana, into the vector pET30a and heterologously...

journal_title：Protein expression and purification

authors： Cao Y,Xian M,Yang J,Xu X,Liu W,Li L

更新日期：2010-02-01 00:00:00

abstract：:Thymosin α1-thymopentin (Tα1-TP5) fusion peptide has been proved to be an immune regulator based on its higher immunoregulatory activity than Tα1 and TP5. To obtain Tα1-TP5 more effectively and economically, Tα1-TP5 was genetically fused to a self-cleaving intein-chitin binding domain tag for purification via chitin b...

journal_title：Protein expression and purification

authors： Li J,Zheng L,Li P,Wang F

更新日期：2012-07-01 00:00:00

abstract：:A specialized vector backbone from the Protein Structure Initiative was used to express full-length human cytochrome b5 as a C-terminal fusion to His8-maltose binding protein in Escherichia coli. The fusion protein could be completely cleaved by tobacco etch virus protease, and a yield of approximately 18 mg of purifi...

journal_title：Protein expression and purification

authors： Sobrado P,Goren MA,James D,Amundson CK,Fox BG

更新日期：2008-04-01 00:00:00

abstract：:The covalent addition of 4-amino-4-deoxy-L-arabinose (L-Ara4N) groups to lipid A, which resides in the outer membranes of bacteria such as Salmonella typhimurium and Escherichia coli, is the final step in the polymyxin-resistance pathway in these organisms. This modification is catalyzed by the inner membrane protein ...

journal_title：Protein expression and purification

authors： Bretscher LE,Morrell MT,Funk AL,Klug CS

更新日期：2006-03-01 00:00:00

abstract：:Escherichia coli oligoribonuclease (EcoORN), encoded by the orn gene, is a 3'-5' exonuclease that degrades short single-stranded oligoribonucleotides to rNMPs in the final step of RNA degradation. The orn gene is essential in E. coli, but not in higher organisms, and close homologues are present in other genomes from ...

journal_title：Protein expression and purification

authors： Young Park A,Elvin CM,Hamdan SM,Wood RJ,Liyou NE,Hamwood TE,Jennings PA,Dixon NE

更新日期：2008-02-01 00:00:00

abstract：:An internal fragment (978 bp) corresponding to the dog zona pellucida glycoprotein-3 (DZP3), excluding the N-terminal signal sequence and the C-terminal transmembrane-like domain, was amplified by polymerase chain reaction from a full-length cDNA clone. The amplified SacI and PstI restricted fragment was cloned in-fra...

journal_title：Protein expression and purification

authors： Santhanam R,Panda AK,Kumar VS,Gupta SK

更新日期：1998-04-01 00:00:00

abstract：:Structural studies of human peptide hormone somatostatin 14 (SS14) require high amounts of isotopically labelled SS14 to be produced. Here we report a method for effective production of isotopically labelled SS14. SS14 was expressed as a fusion protein with thioredoxin in Escherichia coli. Co-expression of a longer po...

journal_title：Protein expression and purification

authors： Nespovitaya N,Barylyuk K,Eichmann C,Zenobi R,Riek R

更新日期：2014-07-01 00:00:00

abstract：:HtrA2 is an apoptosis-activating protein to enhance the apoptotic process by preventing the formation of the IAP-caspase complex, thus freeing caspase to trigger the apoptosis pathway. Here, we presented the full-length sequence of HtrA2 from the black tiger shrimp (PmHtrA2). The full-length PmHtrA2 transcript was 140...

journal_title：Protein expression and purification

authors： Suwannaboon R,Phiwsaiya K,Senapin S,Khunrae P,Rattanarojpong T

更新日期：2013-12-01 00:00:00

abstract：:Human Fas ligand is a medically important membrane glycoprotein that induces the apoptosis of harmful cells. A new secretory expression and purification method was devised for the production of a large amount of recombinant human Fas ligand extracellular domain (hFasLECD) by Pichia pastoris. The expression plasmid con...

journal_title：Protein expression and purification

authors： Muraki M

更新日期：2006-12-01 00:00:00

abstract：:Previously, the lysozyme gene of the Klebsiella phage K11 was partially sequenced in our lab. Using the sequence information the lysozyme gene of the Klebsiella phage K11 was amplified and cloned using the polymerase chain reaction of the pfu DNA polymerase. The nucleotide sequence of phage K11 lysozyme gene was deter...

journal_title：Protein expression and purification

authors： Junn HJ,Youn J,Suh KH,Lee SS

更新日期：2005-07-01 00:00:00