An alternative use of basic pGEX vectors for producing both N- and C-terminal fusion proteins for production and affinity purification of antibodies.

abstract：:Mannan outer chain N-glycan structures are yeast/fungal-specific typically found on secreted and cell wall glycoproteins. Mannan outer chains consist of an alpha-1,6 polymannose backbone attached to a Man(8-10)(GlcNAc)(2) core. The backbone contains branches of alpha-1,2 mannose residues, terminated with alpha-1,3 man...

journal_title：Protein expression and purification

authors： Rodionov D,Romero PA,Berghuis AM,Herscovics A

更新日期：2009-07-01 00:00:00

abstract：:The introduction of an affinity tag offers an attractive approach to isolation of membrane proteins. The type of affinity tag and its positioning in the protein is determined by the desired subsequent experimental uses of the isolated protein. To minimize the risk of interference, membrane proteins may preferentially ...

journal_title：Protein expression and purification

authors： Hamann T,Laursen T,Møller BL

更新日期：2009-11-01 00:00:00

abstract：:In this report, we describe an optimized system for the efficient overexpression, purification, and refolding of secreted bacterial proteins. Candidate secreted proteins were produced recombinantly in Escherichia coli as Tobacco Etch Virus protease-cleavable hexahistidine-c-myc eptiope fusion proteins. Without regard ...

journal_title：Protein expression and purification

authors： Geisbrecht BV,Bouyain S,Pop M

更新日期：2006-03-01 00:00:00

abstract：:We have used Ni(2+)-affinity chromatography as a rapid and efficient method to purify a sensory rhodopsin I (SR-I) derivative containing six consecutive histidine residues at its C-terminus (His-tagged SR-I). The protein was expressed in Halobacterium salinarium by integrating the corresponding gene at the chromosomal...

journal_title：Protein expression and purification

authors： Krebs MP,Spudich EN,Spudich JL

更新日期：1995-12-01 00:00:00

abstract：:A family of restriction enzyme- and ligation-independent cloning vectors has been developed for producing recombinant His-tagged fusion proteins in Escherichia coli. These are based on pURI2 and pURI3 expression vectors which have been previously used for the successful production of recombinant proteins at the millig...

journal_title：Protein expression and purification

authors： Curiel JA,de Las Rivas B,Mancheño JM,Muñoz R

更新日期：2011-03-01 00:00:00

abstract：:2-Deoxy-d-ribose-5-phosphate aldolase (DERA) catalyzes the aldol reaction between two aldehydes and is thought to be a potential biocatalyst for the production of a variety of stereo-specific materials. A gene encoding DERA from the extreme halophilic archaeon, Haloarcula japonica, was overexpressed in Escherichia col...

journal_title：Protein expression and purification

authors： Ohshida T,Hayashi J,Satomura T,Kawakami R,Ohshima T,Sakuraba H

更新日期：2016-10-01 00:00:00

abstract：:A relatively inexpensive yet highly efficient and extremely rapid procedure has been developed for the isolation and purification of estrogen receptor from the goat uterine cytosol. Greater than 1 mg of purified receptor protein could be obtained from 75 g of uterine tissue within a period of < 24 h, following this pr...

journal_title：Protein expression and purification

authors： Zafar A,Thampan RV

更新日期：1993-12-01 00:00:00

abstract：:The high level expression and purification of rat monoamine oxidase B (rMAOB) in the methylotrophic yeast Pichia pastoris is reported. Nearly 100 mg of purified rMAOB is obtained from 130 g (wet weight) of cells (0.5 L of culture). The MALDI-TOF mass spectrum of the purified protein shows a single species with a molec...

journal_title：Protein expression and purification

authors： Upadhyay AK,Edmondson DE

更新日期：2008-06-01 00:00:00

abstract：:(S)-Hydroxynitrile lyase (Hnl) from the tropical rubber tree Hevea brasiliensis catalyzes the formation of (S)-cyanohydrins from hydrocyanic acid and aldehydes or ketones. This enzyme accepts aliphatic, aromatic, and heterocyclic carbonyl compounds as substrates and is therefore considered a potent biocatalyst for the...

journal_title：Protein expression and purification

authors： Hasslacher M,Schall M,Hayn M,Bona R,Rumbold K,Lückl J,Griengl H,Kohlwein SD,Schwab H

更新日期：1997-10-01 00:00:00

abstract：:An unknown d-tagatose 3-epimerase (DTE) containing a IoIE domain was identified and cloned from Escherichia coli. This gene was subcloned into the prokaryotic expression vector pET-15b, and induced by IPTG in E. coli BL21 expression system. Through His-select gel column purification and fast-protein liquid chromatogra...

journal_title：Protein expression and purification

authors： He X,Zhou X,Yang Z,Xu L,Yu Y,Jia L,Li G

更新日期：2015-10-01 00:00:00

abstract：:The supply of many valuable proteins that have potential clinical or industrial use is often limited by their low natural availability. With the modern advances in genomics, proteomics and bioinformatics, the number of proteins being produced using recombinant techniques is exponentially increasing and seems to guaran...

journal_title：Protein expression and purification

authors： Papaneophytou CP,Kontopidis G

更新日期：2014-02-01 00:00:00

abstract：:The type I antifreeze proteins are simple amphipathic helical proteins found in abundance in polar fish species, where they act to prevent freezing of internal fluids by a mechanism of noncolligative freezing point depression. Large-scale production of these proteins for research and biotechnological purposes has been...

journal_title：Protein expression and purification

authors： Solomon RG,Appels R

更新日期：1999-06-01 00:00:00

abstract：:The high specific lysyl endopeptidase (Lys-C; EC 3.4.21.50) is often used for the initial fragmentation of polypeptide chains during protein sequence analysis. However, due to its specificity it could be a useful tool for the production of tailor-made protein hydrolysates with for example bioactive or techno functiona...

journal_title：Protein expression and purification

authors： Stressler T,Eisele T,Meyer S,Wangler J,Hug T,Lutz-Wahl S,Fischer L

更新日期：2016-02-01 00:00:00

abstract：:The cyclin-dependent kinase-activating kinase (CAK) catalyzes the phosphorylation of the cyclin-dependent protein kinases (CDKs) on a threonine residue (Thr160 in human CDK2). The reaction is an obligatory step in the activation of the CDKs. In higher eukaryotes, the CAK complex has been characterized in two forms. Th...

journal_title：Protein expression and purification

authors： Lawrie AM,Tito P,Hernandez H,Brown NR,Robinson CV,Endicott JA,Noble ME,Johnson LN

更新日期：2001-11-01 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：:Bikunin is a proteoglycan exhibiting broad-spectrum inhibitory activity against serine proteases and could potentially suppress tumor cell invasion and metastasis. Here, we have successfully expressed recombinant human bikunin (rh-bikunin) in the methylotrophic yeast Pichia pastoris and established the purification pr...

journal_title：Protein expression and purification

authors： Jian-qiu W,Feng-qin Y,Dou-dou W,Lei B,Nan S,Cong-yan L,Ting Z,Wei-qun Y

更新日期：2008-08-01 00:00:00

abstract：:The alpha-subunit of eukaryotic initiation factor eIF2 (eIF2alpha) plays an important role in the regulation of mRNA translation through modulation of the interaction of eIF2 and a second initiation factor, eIF2B. The interaction of the two proteins is regulated in vivo by phosphorylation of eIF2alpha at Ser51. In the...

journal_title：Protein expression and purification

authors： Kimball SR,Horetsky RL,Jagus R,Jefferson LS

更新日期：1998-04-01 00:00:00

abstract：:The guanine nucleotide exchange factor EF-1beta gene from the thermoacidophilic archaeon Sulfolobus solfataricus (SsEF-1beta) was amplified by PCR and cloned into the pT7-7 expression vector. One of four selected clones harbored the T160C nucleotide substitution leading to the Y54H amino acid change in a hydrophobic r...

journal_title：Protein expression and purification

authors： Ianniciello G,Masullo M,Raimo G,Arcari P,Bocchini V

更新日期：1998-02-01 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

abstract：:Mutant analogues of recombinant human immune interferon (IFN-gamma) with higher stability and biological activity were prepared. Depending on the analogue, protein structure modification might involve introduction of an intramonomer disulfide bond (through replacements of Glu7Cys and Ser69Cys), C-terminal shortening b...

journal_title：Protein expression and purification

authors： Pechenov SE,Tikhonov RV,Shingarova LN,Korobko VG,Yakimov SA,Klyushnichenko VE,Babajantz AA,Beliaev DL,Kuznetzov VP,Shvetz VI,Wulfson AN

更新日期：2002-03-01 00:00:00

abstract：:Disulfide bonds are normally formed after a polypeptide has been exported from the reducing environment of the cytoplasm into a more oxidizing compartment, such as the bacterial periplasm. Recently, we showed that in Escherichia coli trxB gor mutants, in which the reduction of thioredoxin and glutathione is impaired, ...

journal_title：Protein expression and purification

authors： Levy R,Weiss R,Chen G,Iverson BL,Georgiou G

更新日期：2001-11-01 00:00:00

abstract：:Streptomyces coelicolor is a soil-dwelling bacterium that undergoes an intricate, saprophytic lifecycle. The bacterium takes up exogenous nucleosides for nucleic acid synthesis or use as carbon and energy sources. However, nucleosides must pass through the membrane with the help of transporters. In the present work, t...

journal_title：Protein expression and purification

authors： Li F,Liang J,Wang W,Zhou X,Deng Z,Wang Z

更新日期：2015-05-01 00:00:00

abstract：:The gene encoding a novel glucoamylase (GlucaM) from the Corallococcus sp. strain EGB was cloned and heterologous expressed in Escherichia coli BL21(DE3), and the enzymatic characterization of recombinant GlucaM (rGlucaM) was determined in the study. The glucaM had an open reading frame of 1938 bp encoding GlucaM of 6...

journal_title：Protein expression and purification

authors： Li Z,Ji K,Dong W,Ye X,Wu J,Zhou J,Wang F,Chen Q,Fu L,Li S,Huang Y,Cui Z

更新日期：2017-01-01 00:00:00

abstract：:A simple and efficient method for hyperexpression in Escherichia coli and purification of capsid protein, p24, of human immunodeficiency virus type 1 (HIV-1) of both B- and C-subtypes is described. DNA-encoding p24 of C-subtype was cloned from C-subtype gag sequence which was obtained by PCR amplification using DNA ex...

journal_title：Protein expression and purification

authors： Gupta S,Arora K,Gupta A,Chaudhary VK

更新日期：2000-08-01 00:00:00

abstract：:Stem cell factor (SCF) known as the c-kit ligand is a two disulfide bridge-containing cytokine in the regulation of the development and function of hematopoietic cell lineages and other cells such as mast cells, germ cells, and melanocytes. The secreted soluble form of SCF exists as noncovalently associated homodimer ...

journal_title：Protein expression and purification

authors： Akuta T,Kikuchi-Ueda T,Imaizumi K,Oshikane H,Nakaki T,Okada Y,Sultana S,Kobayashi K,Kiyokawa N,Ono Y

更新日期：2015-01-01 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 bivalent anti-human anti-T cell immunotoxin A-dmDT390-bisFv(G(4)S) was developed for treatment of T cell leukemia, autoimmune diseases, and tolerance induction for transplantation. The multi-domain structure of the bivalent immunotoxin hinders efficient production in Escherichia coli and most eukaryotes are sensit...

journal_title：Protein expression and purification

authors： Woo JH,Liu YY,Mathias A,Stavrou S,Wang Z,Thompson J,Neville DM Jr

更新日期：2002-07-01 00:00:00

abstract：:Alphavirus nonstructural protein nsP1 possesses distinct methyltransferase (MTase) and guanylyltransferase (GTase) activities involved in the capping of viral RNAs. In alphaviruses, the methylation of GTP occurs before RNA transguanylation and nsP1 forms a covalent complex with m(7)GMP unlike the host mRNA guanylyltra...

journal_title：Protein expression and purification

authors： Tomar S,Narwal M,Harms E,Smith JL,Kuhn RJ

更新日期：2011-10-01 00:00:00

abstract：:The XRCC1 DNA repair protein contains two regions of approximately 100 amino acids each that share homology with the BRCT (BRCA1 carboxyl terminus) domain superfamily. These two regions of XRCC1 have been shown to interact independently with DNA ligase III and poly(ADP-ribose)polymerase as part of a mechanism involved...

journal_title：Protein expression and purification

authors： Thornton K,Forstner M,Shen MR,West MG,Rupp B,Thelen MP

更新日期：1999-07-01 00:00:00

abstract：:Pigment epithelium-derived factor (PEDF) is a neurotrophic protein and a member of the serine protease inhibitor superfamily. Here we describe the identification of PEDF in bovine eyes and optimization of its purification from this natural source. We have developed a polyclonal antibody to recombinant human PEDF, Ab-r...

journal_title：Protein expression and purification

authors： Wu YQ,Notario V,Chader GJ,Becerra SP

更新日期：1995-08-01 00:00:00