abstract：:The Staphylococcus simulans gene encoding lysostaphin has been PCR amplified from pRG5 recombinant plasmid (ATCC 67076) and cloned into Escherichia coli expression pTYB12 vector (IMPACT-CN System, New England BioLabs) which allows the overexpression of a target protein as a fusion to a self-cleavable affinity tag. The...

journal_title：Protein expression and purification

authors： Szweda P,Pladzyk R,Kotlowski R,Kur J

更新日期：2001-08-01 00:00:00

abstract：:Proteomics has become a major focus as researchers attempt to understand the vast amount of genomic information. Protein complexity makes identifying and understanding gene function inherently difficult. The challenge of studying proteins in a global way is driving the development of new technologies for systematic an...

journal_title：Protein expression and purification

authors： Lesley SA

更新日期：2001-07-01 00:00:00

abstract：:Human prolactin was expressed in insect culture cells by recombinant baculoviruses carrying prolactin gene cDNA placed under the transcriptional control of polyhedrin gene promoter of Autographa californica nuclear polyhedrosis virus. Preliminary results of recombinant human prolactin expression as extracellular as we...

journal_title：Protein expression and purification

authors： Strokovskaya L,Bartoszewicz Z,Szolajska E,Kikhno I,Solomko A,Michalik J

更新日期：2001-07-01 00:00:00

abstract：:CD23, a 45-kDa type II membrane glycoprotein present on B cells, monocytes, and other human immune cells, is a low-affinity receptor for IgE. The extracellular region of the membrane-bound human CD23 is processed into at least four soluble (s) CD23 forms, with apparent molecular masses of 37, 33, 29, and 25 kDa. High ...

journal_title：Protein expression and purification

authors： Khandekar SS,Mayer RJ,Cusimano DM,Katchur SR,Appelbaum ER

更新日期：2001-07-01 00:00:00

abstract：:Leptin is a 16-kDa nonglycosylated hormone that is produced in mature adipocytes and which acts primarily in the hypothalamus to reduce food intake and body weight. While the rat is a representative laboratory animal model in obesity research, so far recombinant rat leptin was not available. In the present study, rat ...

journal_title：Protein expression and purification

authors： Park JH,Lee HH,Na SY,Ju SK,Lee YJ,Lee MK,Kim KL

更新日期：2001-06-01 00:00:00

abstract：:The Plasmodium falciparum cysteine protease falcipain-2 is a potential new target for antimalarial chemotherapy. In order to obtain large quantities of active falcipain-2 for biochemical and structural analysis, a systematic assessment of optimal parameters for the expression and refolding of the protease was carried ...

journal_title：Protein expression and purification

authors： Sijwali PS,Brinen LS,Rosenthal PJ

更新日期：2001-06-01 00:00:00

abstract：:Stromal-cell-derived factor-1 (SDF-1alpha) is an 8-kDa chemokine that is constitutively expressed in bone-marrow-derived stromal cells and has been identified as a ligand for the CXCR4 receptor. We produced the chemokine recombinantly as methionine-SDF-1alpha in Escherichia coli without the leader peptide sequence. Th...

journal_title：Protein expression and purification

authors： Holmes WD,Consler TG,Dallas WS,Rocque WJ,Willard DH

更新日期：2001-04-01 00:00:00

abstract：:We have developed a family of cloning vectors that direct expression of fusion proteins that mimic aggregated immunoglobulin (IgG) (AIG) and immune complex function with respect to their interactions with FcgammaR and that allow for the inclusion and targeting of a second protein domain to cells expressing FcgammaR. T...

journal_title：Protein expression and purification

authors： White DM,Jensen MA,Shi X,Qu Zx,Arnason BG

更新日期：2001-04-01 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：:Protein expression using the secretory pathway in Saccharomyces cerevisiae can lead to high amounts of overexpressed and secreted proteins in culture supernatants in a short period of time. These post-translational modified expression products can be purified up to >90% in a single step. The overexpression and secreti...

journal_title：Protein expression and purification

authors： Schuster M,Wasserbauer E,Aversa G,Jungbauer A

更新日期：2001-02-01 00:00:00

abstract：:Cyclohexanone monooxygenase (CMO) is a soluble flavoenzyme originally isolated from Acinetobacter spp. which carries out Baeyer-Villiger reactions with cyclic ketone substrates. In the present study we cloned the Acinetobacter CMO gene and modified it for facile purification from heterologous expression systems by inc...

journal_title：Protein expression and purification

authors： Cheesman MJ,Kneller MB,Kelly EJ,Thompson SJ,Yeung CK,Eaton DL,Rettie AE

更新日期：2001-02-01 00:00:00

abstract：:Hydrophobic interaction high-performance liquid chromatography (HIC-HPLC) was utilized for the separation of native human antithrombin (AT) and a partially denaturated form of AT, known as the latent form (L-AT). The AT used in this study is commercially available (Atenativ, Pharmacia & Upjohn, Sweden) and contains al...

journal_title：Protein expression and purification

authors： Karlsson G,Winge S

更新日期：2001-02-01 00:00:00

abstract：:To facilitate studies of multicomponent protein complexes, I have developed an Escherichia coli expression system which coexpresses up to four polypeptides from a single plasmid. The modular nature of the system enables efficient subcloning of a gene into each of the 4 cassettes in the polycistronic expression vector....

journal_title：Protein expression and purification

authors： Tan S

更新日期：2001-02-01 00:00:00

abstract：:Wild-type and deglycosylated forms of human prostate-specific antigen were expressed in Chinese hamster ovary (CHO) cells as zymogens. ProPSA was collected from conditioned medium and purified using a single cation-exchange chromatographic step for the deglycosylated form and cation-exchange followed by gel filtration...

journal_title：Protein expression and purification

authors： Bowman KK,Clark J,Yu L,Mortara K,Radika K,Wang J,Zhan H

更新日期：2000-12-01 00:00:00

abstract：:A mammalian expression vector with features optimized for simple expression and purification of secreted proteins has been developed. This vector was constructed to facilitate X-ray crystallographic studies of cysteine-rich glycoproteins that are difficult to express by other means. Proteins expressed with this vector...

journal_title：Protein expression and purification

authors： Leahy DJ,Dann CE 3rd,Longo P,Perman B,Ramyar KX

更新日期：2000-12-01 00:00:00

abstract：:An alpha-amylase has been purified from the thermophilic fungus Scytalidium thermophilum. A ninefold purification was achieved in a single step using fluidized bed chromatography wherein alginate was used as the affinity matrix. There are at least two isoenzymes as shown by concanavalin A (Con A)-agarose column chroma...

journal_title：Protein expression and purification

authors： Roy I,Sastry MS,Johri BN,Gupta MN

更新日期：2000-11-01 00:00:00

abstract：:We examined the feasibility of high-level production of recombinant human prolactin, a multifunctional protein hormone, in insect cells using a baculovirus expression system. The human prolactin cDNA with and without the secretory signal sequence was cloned into pFastBac1 baculovirus vector under the control of polyhe...

journal_title：Protein expression and purification

authors： Das T,Johns PW,Goffin V,Kelly P,Kelder B,Kopchick J,Buxton K,Mukerji P

更新日期：2000-11-01 00:00:00

abstract：:Receptor activator of NF-kappaB ligand (RANKL) is a type II transmembrane protein found on osteoblasts which functions as a major determinant of osteoclast differentiation and activation. RANKL mediates bone homeostasis through binding to the cognate ligand on osteoclasts, RANK, and a soluble decoy receptor, osteoprot...

journal_title：Protein expression and purification

authors： Willard D,Chen WJ,Barrett G,Blackburn K,Bynum J,Consler T,Hoffman C,Horne E,Iannone MA,Kadwell S,Parham J,Ellis B

更新日期：2000-10-01 00:00:00

abstract：:The gene coding for phosphoglucomutase (PGM) from Oryctolagus cuniculus (rabbit) has been expressed in Escherichia coli under a T7 expression system with a His-tag. About half of the expressed PGM protein was present in inclusion bodies, but this protein was inactive when solubilized. The protein in the soluble cell f...

journal_title：Protein expression and purification

authors： Chae YK,Markley JL

更新日期：2000-10-01 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：:A recombinant H(C) fragment of botulinum neurotoxin, serotype A (rBoNTA(H(C))), has been successfully expressed in a Mut(+) strain of the methylotrophic yeast Pichia pastoris for use as an antigen in a proposed human vaccine. Fermentation employed glycerol batch, glycerol-fed batch, and methanol-fed batch phases to ac...

journal_title：Protein expression and purification

authors： Potter KJ,Zhang W,Smith LA,Meagher MM

更新日期：2000-08-01 00:00:00

abstract：:Procedures are described for the preparation of highly purified thymidylate synthases from Escherichia coli and Bacillus subtilis. The yields in each case are quite high with about 350 mg of pure protein obtained from 1 liter of cells. Basically all that is required to obtain pure enzyme is an induction step from a hi...

journal_title：Protein expression and purification

authors： Changchien LM,Garibian A,Frasca V,Lobo A,Maley GF,Maley F

更新日期：2000-07-01 00:00:00

abstract：:The superoxide dismutases (EC 1.15.1.1) are a family of enzymes that catalyze the dismutation of superoxide radical anion to dioxygen and hydrogen peroxide. The active site contains a critical metal ion such as manganese, iron, or copper. The copper-containing protein also has one zinc ion bound per subunit. The stand...

journal_title：Protein expression and purification

authors： Sutter B,Bounds PL,Koppenol WH

更新日期：2000-06-01 00:00:00

abstract：:Human granulocyte-macrophage colony-stimulating factor (GM-CSF), a hemopoietic growth factor, was produced and secreted from tobacco cell suspensions. The GM-CSF cDNA was carried by a binary vector under the control of the CaMV 35S promoter and the T7 terminator. In addition, a 5'-nontranslated region from the tobacco...

journal_title：Protein expression and purification

authors： James EA,Wang C,Wang Z,Reeves R,Shin JH,Magnuson NS,Lee JM

更新日期：2000-06-01 00:00:00

abstract：:The TCL1 gene, which is located on chromosome 14, plays a major role in human hematopoietic malignancies and encodes a 14-kDa protein whose function has not been determined. This gene is expressed in pre-B cells, in immature thymocytes, and, at low levels, in activated T cells but not in peripheral mature B cells and ...

journal_title：Protein expression and purification

authors： Du Bois GC,Song SP,Kulikovskaya I,Rothstein JL,Germann MW,Croce CM

更新日期：2000-04-01 00:00:00

abstract：:Acyl-acyl carrier protein synthase (Aas) is widely used to synthesize thioester adducts of fatty acids between 8 and 18 carbons in length enzymatically to the phosphopantetheine group of acyl carrier protein. The enzyme is an 80.6-kDa inner membrane protein that functions in vivo as a 2-acylglycerophosphoethanolamine ...

journal_title：Protein expression and purification

authors： Shanklin J

更新日期：2000-04-01 00:00:00

abstract：:Pokeweed antiviral protein (PAP)-I from the spring leaves of Phytolacca americana is a naturally occurring RNA-depurinating enzyme with broad-spectrum antiviral activity. Interest in PAP is growing due to its use as a potential anti-HIV agent. However, the clinical use of native PAP is limited due to inherent difficul...

journal_title：Protein expression and purification

authors： Rajamohan F,Doumbia SO,Engstrom CR,Pendergras SL,Maher DL,Uckun FM

更新日期：2000-03-01 00:00:00

abstract：:Quinolinate synthetase catalyzes the second step of the de novo biosynthetic pathway of pyridine nucleotide formation. In particular, quinolinate synthetase is involved in the condensation of dihydroxyacetone phosphate and iminoaspartate to form quinolinic acid. To study the mechanism of action, the specificity of the...

journal_title：Protein expression and purification

authors： Ceciliani F,Caramori T,Ronchi S,Tedeschi G,Mortarino M,Galizzi A

更新日期：2000-02-01 00:00:00

abstract：:Botulinum neurotoxin light chain (BoNT LC, 50 kDa) is responsible for the zinc endopeptidase activity specific for proteins of neuroexocytosis apparatus. We describe the expression of recombinant type A BoNT LC in Escherichia coli as well as the purification and characterization of the recombinant protein. A high leve...

journal_title：Protein expression and purification

authors： Li L,Singh BR

更新日期：1999-12-01 00:00:00

abstract：:Myrosinases are thioglucosidases that hydrolyze the natural plant products glucosinolates. We have expressed the myrosinase MYR1 from Brassica napus in Saccharomyces cerevisiae. The recombinant myrosinase was enzymatically active which shows that the MYR1, which in the plant is complex bound with myrosinase-binding pr...

journal_title：Protein expression and purification

authors： Chen S,Halkier BA

更新日期：1999-12-01 00:00:00

abstract：:The protooncogenic serine/threonine protein kinase PKB contains an amino-terminal pleckstrin homology (PH) domain which binds phosphatidylinositides. The PH domain, composed of approximately 100 loosely conserved amino acids, is found in many proteins, including kinases, phospholipases C, GTPases, GTPase-activating pr...

journal_title：Protein expression and purification

authors： Ingley E,Hemmings BA

更新日期：1999-11-01 00:00:00

abstract：:The A(280)/A(260) ratio of a purified protein is frequently used as an indication of the purity of the preparation with respect to nucleic acids. We show here that for low-molecular-weight recombinant proteins purified from Escherichia coli, a low A(280)/A(260) ratio can also result from contamination with UDP-linked ...

journal_title：Protein expression and purification

authors： Ram MK,Andrade LJ,Phillips TB,van Schravendijk MR

更新日期：1999-11-01 00:00:00

abstract：:NIP1, the product of the avirulence gene AvrRrs1 from Rhynchosporium secalis, a fungal pathogen of barley, is a small secreted cysteine-rich protein. This protein is essential for the specific recognition of the fungus by host plants carrying the complementary resistance gene Rrs1. Different heterologous expression sy...

journal_title：Protein expression and purification

authors： Gierlich A,van 't Slot KA,Li VM,Marie C,Hermann H,Knogge W

更新日期：1999-10-01 00:00:00

abstract：:Large amounts of soluble fibronectin were easily purified from cryoprecipitated or fresh citrated human blood plasma by a three-step combination of gelatin and heparin-cellufine affinity chromatography. The elution conditions were optimized to obtain a homogeneous fraction on SDS-PAGE and Western blot under reducing c...

journal_title：Protein expression and purification

authors： Poulouin L,Gallet O,Rouahi M,Imhoff JM

更新日期：1999-10-01 00:00:00

abstract：:Achromogenic atypical Aeromonas salmonicida is the causative agent of goldfish ulcer disease. Virulence of this bacterium is associated with the production of a paracrystalline outer membrane A-layer protein. The species-specific structural gene for the monomeric form of A-protein was cloned into a pET-3d plasmid in o...

journal_title：Protein expression and purification

authors： Maurice S,Hädge D,Dekel M,Friedman A,Gertler A,Shoseyov O

更新日期：1999-08-01 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：:A cysteine-rich (approximately 20%), low molecular weight (MW 6 kDa) polypeptide has been isolated from the Korean blood-sucking leech, Hirudo nipponia. From its amino acid composition and N-terminal amino acid sequence analysis, the new protein is similar to granulin (or epithelin), and so it has been named leech gra...

journal_title：Protein expression and purification

authors： Hong SJ,Kang KW

更新日期：1999-07-01 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 gene for the extremely thermophilic and thermostable 5'-methylthioadenosine phosphorylase from the archaeon Sulfolobus solfataricus was expressed at a high level in Escherichia coli thus providing a basis for detailed structural and functional studies of the enzyme. The recombinant enzyme was purified to homogenei...

journal_title：Protein expression and purification

authors： Cacciapuoti G,Fusco S,Caiazzo N,Zappia V,Porcelli M

更新日期：1999-06-01 00:00:00

abstract：:A method for the simplified, reproducible production of both normal and altered versions of human proinsulin has been developed. A polyhistidine/proinsulin fusion protein was expressed using a prokaryotic expression system and partially purified by affinity chromatography. Disulfide bonds within the polypeptide were f...

journal_title：Protein expression and purification

authors： Mackin RB

更新日期：1999-04-01 00:00:00