abstract：:A new method was developed to purify the streptococcal hyaluronate synthase in active form to electrophoretic homogeneity. The method is based on the extraction of protoplast membranes with digitonin and a phase separation into an aqueous and a detergent phase induced by addition of polyethylene glycol 6000 at 0 degre...

journal_title：Protein expression and purification

authors： Prehm S,Nickel V,Prehm P

更新日期：1996-06-01 00:00:00

abstract：:We have cloned, from total human liver RNA, the cDNA encoding apolipoprotein E3 (apoE3). Site-directed mutagenesis was used to obtain the cDNA encoding the apoE4 isoform, a major variant of this apolipoprotein in man. These two cDNAs were subcloned into the procaryotic expression vector pAHRS. A polyhistidine tag was ...

journal_title：Protein expression and purification

authors： Pillot T,Barbier A,Visvikis A,Lozac'h K,Rosseneu M,Vandekerckhove J,Siest G

更新日期：1996-06-01 00:00:00

abstract：:Human pancreatic ribonuclease (HP-RNase) has considerable promise as a therapeutic agent. Structure-function analyses of HP-RNase have been impeded by the difficulty of obtaining the enzyme from its host. Here, a gene encoding HP-RNase was designed, synthesized, and inserted into two expression vectors that then direc...

journal_title：Protein expression and purification

authors： Ribó M,delCardayré SB,Raines RT,de Llorens R,Cuchillo CM

更新日期：1996-05-01 00:00:00

abstract：:Hepatocyte growth factor (HGF) is a multifunctional protein expressed in a variety of cell types and tissues. Here we describe a novel one-step method to separate and identify HGF, based on a unique interaction between HGF and Cu(II). Conditioned medium (CM) from mouse 3T3-L1 adipocytes which contains HGF or purified ...

journal_title：Protein expression and purification

authors： Rahimi N,Etchells S,Elliott B

更新日期：1996-05-01 00:00:00

abstract：:Anthrax toxin consists of three separate proteins, protective antigen (PA), lethal factor (LF), and edema factor (EF). PA binds to the receptor on mammalian cells and facilitates translocation of EF or LF into the cytosol. PA is the primary component of several anthrax vaccines. In this study we expressed and purified...

journal_title：Protein expression and purification

authors： Sharma M,Swain PK,Chopra AP,Chaudhary VK,Singh Y

更新日期：1996-02-01 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：:Rat granulocyte macrophage colony-stimulating factor (rGM-CSF) cDNA was amplified and cloned, and recombinant-rGM-CSF (R-rGM-CSF) was expressed and isolated from Escherichia coli. The synthesis of R-rGM-CSF was directed by a modified, inducible maltose binding protein (MBP) gene fusion expression vector, pMTR-23, and ...

journal_title：Protein expression and purification

authors： Holowachuk EW,Ruhoff MS

更新日期：1995-10-01 00:00:00

abstract：:Ricin A chain (RTA) mutants which had been modified by the addition of three lysine residues, three lysines and an alanine, or six histidine residues at the carboxyl terminus were expressed in Escherichia coli. The recombinant proteins were purified to homogeneity by ion-exchange chromatography on CM-Sepharose CL-6B. ...

journal_title：Protein expression and purification

authors： Simpson JC,Roberts LM,Lord JM

更新日期：1995-10-01 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

abstract：:Ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit epsilon N-methyltransferase (Protein methylase III, Rubisco LSMT, EC 2.1.1.43) catalyzes methylation of the epsilon-amino group of Lys-14 in the large subunit of Rubisco. In this paper, an affinity purification procedure for pea (Pisum sativum L. cv Laxton'...

journal_title：Protein expression and purification

authors： Wang P,Royer M,Houtz RL

更新日期：1995-08-01 00:00:00

abstract：:A cDNA clone, lambda GTHP1del, encoding glutathione transferase (GST) P1-1, was isolated from a human K562 erythroleukemia cell line cDNA library. The coding sequence was lacking the codons for the N-terminal 34 amino acids. A DNA segment was designed in order to obtain the missing portion and a structure representing...

journal_title：Protein expression and purification

authors： Kolm RH,Stenberg G,Widersten M,Mannervik B

更新日期：1995-06-01 00:00:00

abstract：:The basidiomycete Phanerochaete chrysosporium produces several isoforms of lignin peroxidase, which catalyzes the oxidative depolymerization of lignin To date, ion-exchange chromatography and preparative isoelectric focusing (IEF) have been commonly used for isolation of lignin peroxidase isoenzymes. In this work we h...

journal_title：Protein expression and purification

authors： Ollikka P,Leppänen VM,Anttila T,Suominen I

更新日期：1995-06-01 00:00:00

abstract：:Bovine annexin IV, a Ca(2+)-dependent, membrane-binding protein, was expressed in E. coli using four different prokaryotic expression vector systems. An annexin IV cDNA was mutated in the 5' noncoding region to introduce an NcoI restriction site at the translation initiation site. The coding sequence was then excised ...

journal_title：Protein expression and purification

authors： Nelson MR,Creutz CE

更新日期：1995-04-01 00:00:00

abstract：:Cell extracts of Clostridium kluyveri grown on ethanol plus succinate contained a NAD(H) dependent 4-hydroxybutanoate dehydrogenase (EC 1.1.1.61) at 66 U/mg. This enzyme was purified 42-fold under anaerobic conditions to homogeneity. Heat treatment, ion exchange chromatography on DEAE-cellulose, nondenaturing polyacry...

journal_title：Protein expression and purification

authors： Wolff RA,Kenealy WR

更新日期：1995-04-01 00:00:00

abstract：:Recombinant human interleukin-5 (rhIL-5) was expressed in baculovirus-infected insect cells and purified to homogeneity from the culture medium in a single chromatographic step. Beginning with a cDNA encoding the full-length precursor form of human IL-5, including the authentic secretory leader sequence, recombinant b...

journal_title：Protein expression and purification

authors： Brown PM,Scheid MP,O'Neill GP,Tagari PC,Nicholson DW

更新日期：1995-02-01 00:00:00

abstract：:The present report documents an improved enzyme assay method for the mammalian L-alanine:4,5-dioxovalerate transaminase which is of significant utility in work with crude tissue homogenates, cell cultures, or purified enzyme preparations. We also describe a new and rapid purification procedure for this enzyme from rat...

journal_title：Protein expression and purification

authors： Tyagi RK,Datta K

更新日期：1994-12-01 00:00:00

abstract：:The assembly pathway of the insect cell Spodoptera frugiperda (Sf-9) was engineered to include expression of the murine chaperone immunoglobulin heavy chain binding protein (BiP) using the baculovirus vector. The impact of BiP coexpression on the production and secretion of functional and soluble recombinant immunoglo...

journal_title：Protein expression and purification

authors： Hsu TA,Eiden JJ,Bourgarel P,Meo T,Betenbaugh MJ

更新日期：1994-12-01 00:00:00

abstract：:L-Threonine dehydrogenase was purified 10,000-fold to a specific activity approximately 300 mumol.min-1.mg-1 protein from porcine liver mitochondria. Purification to apparent homogeneity was achieved by sequential chromatography on DEAE Sepharose FF, Affi-Gel Blue, Sephacryl S-200, Matrex Gel Red A, and Matrex Gel Gre...

journal_title：Protein expression and purification

authors： Kao YC,Davis L

更新日期：1994-10-01 00:00:00

abstract：:Human GRO alpha, GRO beta, and GRO gamma are neutrophil chemoattractants structurally related to IL-8 and compete with IL-8 for binding to IL-8 receptors on neutrophils. These proteins are part of a large superfamily of chemotactic cytokines, the "chemokines," members of which share striking structural similarities. W...

journal_title：Protein expression and purification

authors： Zagorski J,DeLarco JE

更新日期：1994-08-01 00:00:00

abstract：:A maize dehydrin with an apparent molecular weight of 20 kDa was purified from whole kernels of maize inbred line G50. Kernels were ground in a seed mill, stirred overnight in extraction buffer, and centrifuged to extract soluble proteins. The sample was heated to 89 degrees C and centrifuged to remove heat-insoluble ...

journal_title：Protein expression and purification

authors： Ceccardi TL,Meyer NC,Close TJ

更新日期：1994-06-01 00:00:00

abstract：:A method was developed for the affinity purification of human complement properdin. The preparation is part of an integrated scheme in which over 20 human plasma proteins can be recovered in a highly purified form. The yield of properdin was 5.9 mg from 3 liters of plasma, amounting to a 28% recovery. The crucial step...

journal_title：Protein expression and purification

authors： DiScipio RG

更新日期：1994-04-01 00:00:00

abstract：:Escherichia coli cells were transformed with an expression vector constructed by inserting a DNA fragment encoding a Kazal-type trypsin inhibitor from mouse seminal vesicle into pGEX-2. The cloned cells were able to produce a high yield of a chimeric polypeptide made by fusing the trypsin inhibitor to glutathione S-tr...

journal_title：Protein expression and purification

authors： Lai ML,Li SH,Chen YH

更新日期：1994-02-01 00:00:00

abstract：:Azotobacter vinelandii has recently been used for a variety of genetic experiments which take advantage of its facile transformation system and its high-frequency homologous recombination. One gene that has been cloned and sequenced is the fdxA gene that encodes a small Fe-S protein called A. vinelandii ferredoxin I (...

journal_title：Protein expression and purification

authors： Vázquez A,Shen B,Negaard K,Iismaa S,Burgess B

更新日期：1994-02-01 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：:A baculovirus vector system that expresses cloned DNA sequences as glutathione S-transferase fusion proteins was developed. This system was used to express and purify the lymphocyte-specific tyrosine kinase p56lck. This recombinant p56lck was purified to homogeneity in a single chromatography step using glutathione re...

journal_title：Protein expression and purification

authors： Spana C,O'Rourke EC,Bolen JB,Fargnoli J

更新日期：1993-10-01 00:00:00

abstract：:Haloalkane dehalogenase from Xanthobacter autotrophicus was efficiently expressed in Escherichia coli BL21 (DE3) and E. coli JM101. After introduction of restriction sites by PCR the haloalkane dehalogenase gene (dhlA) was translationally fused behind the T7 (phi 10), trc, and tac promoters. This resulted in expressio...

journal_title：Protein expression and purification

authors： Schanstra JP,Rink R,Pries F,Janssen DB

更新日期：1993-10-01 00:00:00

abstract：:The purification of biologically active human protein synthesis initiation factor 4 alpha, eIF-4 alpha, overexpressed in Escherichia coli, is complicated by its localization in insoluble inclusion bodies, as well as its possession of four cysteines. Two of these cysteines have been reported to be reduced in the native...

journal_title：Protein expression and purification

authors： Stern BD,Wilson M,Jagus R

更新日期：1993-08-01 00:00:00

abstract：:An efficient expression system for a low-molecular mass xylanase in Escherichia coli has been developed. A gene encoding the mature Bacillus circulans (Bc) xylanase was designed to imitate the frequency of degenerate codons used in E. coli. Appropriate degenerate codons were used to create multiple unique restriction ...

journal_title：Protein expression and purification

authors： Sung WL,Luk CK,Zahab DM,Wakarchuk W

更新日期：1993-06-01 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：:The cDNA for the alpha i1 protein that had undergone site-directed mutagenesis to change glycine-2 to alanine was ligated into a baculovirus transfer vector. A recombinant virus was obtained by transfecting Sf9 cells with both the wild-type baculovirus DNA and the transfer vector and screening for recombinant plaques....

journal_title：Protein expression and purification

authors： Jones TL,Woodard C,Spiegel AM

更新日期：1993-02-01 00:00:00

abstract：:Lectins are a class of proteins with specific carbohydrate-binding properties found in a wide variety of plants and animals. Gramineae lectins are presumably defense-related proteins in plants that exert their effect by binding to N-acetylglucosamine. Barley lectin is a vacuolar protein synthesized with an amino-termi...

journal_title：Protein expression and purification

authors： Schroeder MR,Raikhel NV

更新日期：1992-12-01 00:00:00

abstract：:General procedures for the rapid, large-scale purification of recombinant Lactobacillus casei thymidylate synthase and its mutants have been established. An effective method employs sequential phosphocellulose and hydroxylapatite chromatography. Crude cell extracts are directly applied to phosphocellulose, and the enz...

journal_title：Protein expression and purification

authors： Kealey JT,Santi DV

更新日期：1992-10-01 00:00:00

abstract：:Streptococcal protein G has been used extensively for the purification of antibodies using the interaction of the Fc region with protein G. Many antibodies also interact with protein G through a low-affinity binding site for the Fab region. The exploitation of this low-affinity interaction for the purification of Fab'...

journal_title：Protein expression and purification

authors： Proudfoot KA,Torrance C,Lawson AD,King DJ

更新日期：1992-10-01 00:00:00

abstract：:To study the subunit structure and the active site of human immunodeficiency virus reverse transcriptase (RT), the enzyme was expressed in E. coli and purified to homogeneity in large quantities. The recombinant enzyme consists of two major polypeptides of 66,000 and 53,000 Da in equimolar amounts and a minor species ...

journal_title：Protein expression and purification

authors： Fischer M,Lifshitz R,Katz T,Liefer I,Ben-Artzi H,Gorecki M,Panet A,Zeelon E

更新日期：1992-08-01 00:00:00

abstract：:Separation of copurifying protease activity from recombinant human Müllerian inhibiting substance (rhMIS) bound to a monoclonal antibody immunoaffinity column by a high-salt wash results in cleaner preparations of rhMIS resistant to cleavage upon storage. In addition, an inhibitor of rhMIS antiproliferative activity i...

journal_title：Protein expression and purification

authors： Ragin RC,Donahoe PK,Kenneally MK,Ahmad MF,MacLaughlin DT

更新日期：1992-06-01 00:00:00

abstract：:Phenylalanine hydroxylase, important in phenylalanine metabolism in mammals, is regulated through short-term (activation) and long-term (induction) mechanisms. To help elucidate the structure-function relationships involved in the activation of this enzyme, we have isolated and characterized full-length cDNA clones to...

journal_title：Protein expression and purification

authors： Citron BA,Davis MD,Kaufman S

更新日期：1992-04-01 00:00:00

abstract：:A procedure for the purification of the rat liver microsomal carnitine octanoyltransferase (COT) that catalyzes the reversible formation of medium-chain and long-chain acylcarnitines from acyl-coenzyme A is described. The K0.5 for L-carnitine is 0.6 mM and the K0.5 for both decanoyl-CoA and palmitoyl-CoA is 0.6 microM...

journal_title：Protein expression and purification

authors： Murthy MS,Bieber LL

更新日期：1992-02-01 00:00:00

abstract：:Strategies for the expression of precursors of eukaryotic secreted proteins as part of fused proteins in Escherichia coli have been explored. A fusion protein with beta-galactosidase at the N-terminal end and honeybee prepromelittin at the C-terminal end (beta-gal-pM) was expressed in low amounts as a cleaved polypept...

journal_title：Protein expression and purification

authors： He M,Adcock I,Chapman D,Lucy J,Austen B

更新日期：1991-10-01 00:00:00

abstract：:We have constructed a plasmid, pAHOB1, with a 482-b AluI fragment containing the Escherichia coli glutaredoxin gene (grx) cloned under lambda PL promoter control. Growth of E. coli N4830/pAHOB1 cells at 30 degrees C followed by heat induction at 40 degrees C for 5 h resulted in expression of glutaredoxin as 20% of the...

journal_title：Protein expression and purification

authors： Björnberg O,Holmgren A

更新日期：1991-08-01 00:00:00

abstract：:A fairly rapid and improved method for producing large amounts of highly pure apoaequorin, the apoprotein of aequorin which emits light on binding Ca2+, is described. The method consists of fusing the gene of the outer membrane protein A (ompA) secretion signal peptide of Escherichia coli to the apoaequorin gene and e...

journal_title：Protein expression and purification

authors： Inouye S,Zenno S,Sakaki Y,Tsuji FI

更新日期：1991-04-01 00:00:00