abstract：:A high-expression plasmid, pKSGHE3-1, containing the salt-tolerant glutaminase (EC 3.5.1.2) from marine bacterium Micrococcus luteus K-3 was constructed. pKSGHE3-1 was made by inserting the DNA fragment (1.43 kb) containing the structural gene synthesized by polymerase chain reaction into the downstream region of the ...

journal_title：Protein expression and purification

authors： Nandakumar R,Wakayama M,Nagano Y,Kawamura T,Sakai K,Moriguchi M

更新日期：1999-03-01 00:00:00

abstract：:Recombinant murine MRP14 (mMRP14) was produced in Escherichia coli using the pGEX expression system. The mass of fusion protein, by electrospray ionization-mass spectrometry (ESI/MS), was 39,213 Da which compares well with the theoretical mass (39,210.4 Da). Thrombin digestion of fusion protein was expected at a clone...

journal_title：Protein expression and purification

authors： Raftery MJ,Collinson L,Geczy CL

更新日期：1999-03-01 00:00:00

abstract：:Ketol-acid reductoisomerase (EC 1.1.1.86) catalyzes the conversion of 2-aceto-2-hydroxyacids to 2-keto-3-hydroxyacids and their subsequent reduction by NADPH to 2,3-dihydroxyacids. The gene encoding the Escherichia coli enzyme was cloned and expressed as a hexahistidine-tagged fusion protein and the recombinant enzyme...

journal_title：Protein expression and purification

authors： Hill CM,Duggleby RG

更新日期：1999-02-01 00:00:00

abstract：:The kinetic locking-on strategy improves the selectivity of protein purification procedures based on immobilized cofactor derivatives through use of enzyme-specific substrate analogues in irrigants to promote biospecific adsorption. This paper describes the development and application of this strategy to the one-chrom...

journal_title：Protein expression and purification

authors： O'flaherty M,McMahon M,Mulcahy P

更新日期：1999-02-01 00:00:00

abstract：:For affinity-chromatography-based purification of proteins that are prone to abnormal termination of translation or that may not be modified at their N-termini, affinity tags are needed which can be fused to the C-terminus. In this publication we describe that maltose binding protein (MBP) fused to the C-terminus of t...

journal_title：Protein expression and purification

authors： Hennig L,Schäfer E

更新日期：1998-12-01 00:00:00

abstract：:The heat shock protein hsp60 plays a functional role in insulin-dependent diabetes mellitus. The hsp60 epitope p277 (aa 437-aa 460) is effective in vaccinating mice against diabetes. A synthetic peptide gene (p277) that encodes the human hsp60 epitope was cloned to the 3' end of the cellulose-binding domain gene (cbd)...

journal_title：Protein expression and purification

authors： Shpigel E,Elias D,Cohen IR,Shoseyov O

更新日期：1998-11-01 00:00:00

abstract：:Homocysteine alpha,gamma-lyase from the anaerobic protozoan parasite Trichomonas vaginalis has been cloned from genomic DNA using PCR methods and expressed in Escherichia coli with a vector containing the T7 promoter. The recombinant homocysteine alpha,gamma-lyase (rHCYase) is expressed as the major protein in the hos...

journal_title：Protein expression and purification

authors： Han Q,Lenz M,Tan Y,Xu M,Sun X,Tan X,Tan X,Tang L,Miljkovic D,Hoffman RM

更新日期：1998-11-01 00:00:00

abstract：:Previously, we have shown that the entire extracellular domain of the granulocyte-colony stimulating factor receptor (sG-CSFr) produced in Chinese hamster ovary (CHO) cells forms a stable complex with its ligand G-CSF, at a stoichiometry of 2:2. A truncated receptor molecule consisting of the cytokine receptor homolog...

journal_title：Protein expression and purification

authors： Horan TP,Simonet L,Jacobsen R,Mann M,Haniu M,Wen J,Arakawa T,Kuwamoto M,Martin F

更新日期：1998-10-01 00:00:00

abstract：:Soybean root nodules possess a developmentally regulated acid phosphatase (ACP) that exhibits the highest specificity for purine 5'-nucleoside monophosphates. The enzyme is a glycosylated dimer of 28- and 31-kDa subunits, which appear to be products of the same gene but differ in posttranslational modifications. In or...

journal_title：Protein expression and purification

authors： Penheiter AR,Klucas RV,Sarath G

更新日期：1998-10-01 00:00:00

abstract：:A recombinant Hc fragment of botulinum neurotoxin, serotype B (rBoNTB(Hc)), has been successfully expressed in a Mut+ strain of the methylotrophic yeast Pichia pastoris for use as an antigen in a proposed human vaccine. The fermentation process consisted of batch phase on glycerol, followed by glycerol and methanol fe...

journal_title：Protein expression and purification

authors： Potter KJ,Bevins MA,Vassilieva EV,Chiruvolu VR,Smith T,Smith LA,Meagher MM

更新日期：1998-08-01 00:00:00

abstract：:A rabbit liver cDNA library in phage lambdagt10 was screened using the coding cDNA for human cytosolic serine hydroxymethyltransferase. A clone of 1754 bp was isolated and the nucleotide sequence showed an open reading frame of 1455 bp, which coded for rabbit cytosolic serine hydroxymethyltransferase and was flanked b...

journal_title：Protein expression and purification

authors： di Salvo ML,Delle Fratte S,De Biase D,Bossa F,Schirch V

更新日期：1998-07-01 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：:The UDP-N-acetylmuramoyl-L-alanine:D-glutamate ligase from Escherichia coli, an enzyme involved in the biosynthesis of the bacterial peptidoglycan monomer unit, was overproduced and purified to homogeneity on a large scale, yielding 4 mg of protein per liter of bacterial culture. Crystals of the complex with the subst...

journal_title：Protein expression and purification

authors： Auger G,Martin L,Bertrand J,Ferrari P,Fanchon E,Vaganay S,Pétillot Y,van Heijenoort J,Blanot D,Dideberg O

更新日期：1998-06-01 00:00:00

abstract：:Taxadiene synthase catalyzes the conversion of the universal precursor of diterpenoids, geranylgeranyl diphosphate, to taxadiene, a key intermediate in Taxol (paclitaxel) biosynthesis. The gene encoding taxadiene synthase was cloned recently. Here we report a method for the heterologous overexpression of cDNA encoding...

journal_title：Protein expression and purification

authors： Huang KX,Huang QL,Wildung MR,Croteau R,Scott AI

更新日期：1998-06-01 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：: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：:Keratinocyte growth factor (KGF) is a member of the fibroblast growth factor family that acts specifically on epithelial cells in a paracrine mode. We employed a mammalian expression system to synthesize recombinant human KGF and isolated two preparations, KGF-a and KGF-b, from medium conditioned by Chinese hamster ov...

journal_title：Protein expression and purification

authors： Hsu YR,Hsu EW,Katta V,Brankow D,Tseng J,Hu S,Morris CF,Kenney WC,Lu HS

更新日期：1998-03-01 00:00:00

abstract：:A novel and rapid procedure for the isolation of catalase from mouse liver, after prior treatment with the peroxisome proliferator perfluorooctanoic acid was developed using immobilized metal ion affinity chromatography involving chelation with zinc ions. The purification developed is simple, rapid (requiring 3 hours ...

journal_title：Protein expression and purification

authors： Yang Q,DePierre JW

更新日期：1998-03-01 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：:The hepatitis E virus (HEV) capsid antigen has been proposed as a candidate subunit vaccine for the prevention of hepatitis E. The full-length HEV ORF2 protein product is predicted to contain 660 amino acids and to weigh 72,000 daltons. Expression of the HEV ORF2 capsid gene from recombinant baculoviruses in insect ce...

journal_title：Protein expression and purification

authors： Robinson RA,Burgess WH,Emerson SU,Leibowitz RS,Sosnovtseva SA,Tsarev S,Purcell RH

更新日期：1998-02-01 00:00:00

abstract：:During the screening for tyrosine phenol-lyase-producing thermophiles, we isolated an obligatory symbiotic thermophile, Symbiobacterium sp. SC-1, which grew only in coculture with Bacillus sp. SK-1. A gene encoding thermostable tyrosine phenol-lyase (TPL) was cloned from the genomic DNA of the Symbiobacterium sp. SC-1...

journal_title：Protein expression and purification

authors： Lee SG,Hong SP,Choi YH,Chung YJ,Sung MH

更新日期：1997-12-01 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 Escherichia coli expression system that exploits the bacterial alkaline phosphatase (PhoA) signal sequence to translocate recombinant human epidermal growth factor (hEGF) to the periplasm was used to evaluate how changes in the composition and sequence of amino acids near the PhoA-hEGF junction influence the peripl...

journal_title：Protein expression and purification

authors： Campion SR,Elsasser E,Chung R

更新日期：1997-08-01 00:00:00

abstract：:The gene that was inferred to encode the TyrR protein of Haemophilus influenzae Rd was synthesized by polymerase chain reaction and inserted into a T7-based expression vector. Methods were developed to overexpress the TyrR protein of H. influenzae in Escherichia coli and to purify the protein on a large scale. Both in...

journal_title：Protein expression and purification

authors： Zhu Q,Zhao S,Somerville RL

更新日期：1997-07-01 00:00:00

abstract：:The cDNA encoding the 65-kDa subunit of malic enzyme from Ascaris suum was cloned into the bacterial expression vector pKK223-3 and overproduced in Escherichia coli. A protein with a subunit molecular mass of 65,000 was expressed at a level of up to 3% of the total soluble protein in JM109, as judged by SDS-PAGE. The ...

journal_title：Protein expression and purification

authors： Chooback L,Karsten WE,Kulkarni G,Nalabolu SR,Harris BG,Cook PF

更新日期：1997-06-01 00:00:00

abstract：:Transcription regulation in the cell occurs in the context of chromatin. It follows that a thorough investigation of the mechanism of transcription regulation must take into account the role of chromatin structure. Through classical and molecular genetic experiments in yeast, great strides have been made in understand...

journal_title：Protein expression and purification

authors： Pilon J,Terrell A,Laybourn PJ

更新日期：1997-06-01 00:00:00

abstract：:Human HEK293 cells that stably express the Epstein Barr nuclear antigen 1 (EBNA1) support the episomal replication of plasmids containing the Epstein Barr virus origin of replication (EBV oriP). A 293EBNA (293E) cell line expressing the human corticotropin-releasing hormone receptor subtype I (CRHR1) from an episomal ...

journal_title：Protein expression and purification

authors： Horlick RA,Sperle K,Breth LA,Reid CC,Shen ES,Robbins AK,Cooke GM,Largent BL

更新日期：1997-04-01 00:00:00

abstract：:Cysteine-rich intestinal protein (CRIP) is a small, 8.5-kDa protein with one double zinc-finger motif called a LIM domain. It is very abundant in intestine and some immune cells in rodents, and expression is influenced by development and the immune response. We have cloned a human CRIP cDNA from human small intestine ...

journal_title：Protein expression and purification

authors： Khoo C,Blanchard RK,Sullivan VK,Cousins RJ

更新日期：1997-04-01 00:00:00

abstract：:The human cyclin H, a protein normally associated with the cyclin-dependent kinase cdk7, was overexpressed in Escherichia coli using a T7 RNA polymerase expression system and further purified to apparent homogeneity. The purified recombinant cyclin H is similar to the endogenous one according to the following criteria...

journal_title：Protein expression and purification

authors： Poterszman A,Andersen G,Busso D,Rossignol M,Egly JM,Thierry JC

更新日期：1997-03-01 00:00:00

abstract：:Chinese hamster dihydrofolate reductase (ch-DHFR) was overexpressed in Escherichia coli DH5 alpha under the transcriptional control of PRPL promoters regulated by temperature-sensitive repressors. The desired recombinant product is soluble and constitutes about 30% of the total soluble proteins of the bacterial cell. ...

journal_title：Protein expression and purification

authors： Fan YX,Wang B,Jing GZ

更新日期：1997-03-01 00:00:00

abstract：:Hydroxypyruvate reductase (HPR), a plant leaf peroxisomal enzyme involved in the glycolate pathway, has been purified in two steps from a crude extract of parsley leaves during the purification of an unrelated ATP-dependent enzyme. HPR, a homogenous side-fraction arising from this purification procedure, was identifie...

journal_title：Protein expression and purification

authors： Julliard JH,Breton-Gilet A

更新日期：1997-02-01 00:00:00

abstract：:NF-kappaB is a pleiotropic transcriptional activator originally identified by its ability to regulate immunoglogulin kappa light chain expression. Purification of this DNA-binding complex demonstrated that NF-kappaB is a heterodimer composed of two subunits, NFKB1 and RelA. Previous studies have shown that truncated v...

journal_title：Protein expression and purification

authors： Coleman TA,Huddleston KA,Ruben SM,Rosen CA,Gentz R

更新日期：1997-02-01 00:00:00

abstract：:Escherichia coli glutamic acid decarboxylase is a pyridoxal phosphate-dependent enzyme that catalyzes the alpha-decarboxylation of glutamate to yield 4-amino-butyrate and CO2. The E. coli chromosome contains two genes encoding for this enzyme, gadA and gadB, which map at distinct loci. Their protein products differ in...

journal_title：Protein expression and purification

authors： De Biase D,Tramonti A,John RA,Bossa F

更新日期：1996-12-01 00:00:00

abstract：:Glycolate oxidase is a flavin-dependent enzyme in the photorespiratory pathway in plants. Here we report the heterologous expression of glycolate oxidase in Escherichia coli and an isolation procedure which results in 4 mg pure protein per gram cell paste in only 1.5 days. This corresponds to a more than 50-fold impro...

journal_title：Protein expression and purification

authors： Stenberg K,Lindqvist Y

更新日期：1996-11-01 00:00:00

abstract：:The hexokinase (ATP;D-hexose 6-phosphotransferase, EC 2.7.1.1) of Schistosoma mansoni has been expressed in Escherichia coli as a fusion protein including an N-terminal polyhistidine tag and enterokinase cleavage site. The enzyme was purified by metal chelate chromatography to > 95% homogeneity, based on analysis by S...

journal_title：Protein expression and purification

authors： Armstrong RL,Wilson JE,Shoemaker CB

更新日期：1996-11-01 00:00:00

abstract：:The entire extracellular domain of the human heat-stable enterotoxin (ST) receptor as well as a truncated N-terminal domain were cloned as glutathione S-transferase fusion proteins and expressed in Escherichia coli. The recombinant fusion proteins were purified from both the cytosol and the inclusion body fractions by...

journal_title：Protein expression and purification

authors： Nandi A,Mathew R,Visweswariah SS

更新日期：1996-09-01 00:00:00

abstract：:Baculovirus expression vectors are used routinely for foreign gene expression and are under intense development as improved biological pesticides. Conventional baculovirus expression vectors are recombinant viruses that can express a foreign gene in insect cells under the control of the polyhedrin promoter, which prov...

journal_title：Protein expression and purification

authors： Jarvis DL,Weinkauf C,Guarino LA

更新日期：1996-09-01 00:00:00

abstract：:We describe a single method to purify milligram amounts of authentic wild-type and mutant HIV-1 and HIV-2 Tat proteins overexpressed in Escherichia coli. This method takes advantage of the highly basic, positively charged RNA binding domain present in both HIV-1 and HIV-2 Tat, which also facilitated purification of HI...

journal_title：Protein expression and purification

authors： Orsini MJ,García-Martínez LF,Mavankal G,Gaynor RB,Debouck CM

更新日期：1996-09-01 00:00:00

abstract：:This paper reports the overproduction and the details of a rapid method to purify active sigmaS monomers from a T7 RNA polymerase-based protein expression system. This 2-day procedure involves solubilizing inclusion bodies in sarkosyl detergent, removal of sarkosyl by dialysis, and a single gel filtration column chrom...

journal_title：Protein expression and purification

authors： Nguyen LH,Burgess RR

更新日期：1996-08-01 00:00:00

abstract：:In order to produce sufficient quantities of fibroblast growth factor-saporin (rFGF-2-SAP) mitotoxin for preclinical evaluation in models of diseases such as cancer and restenosis, we have undertaken the large-scale expression, purification, and characterization of the recombinant molecule. The fusion gene encoding rF...

journal_title：Protein expression and purification

authors： McDonald JR,Ong M,Shen C,Parandoosh Z,Sosnowski B,Bussell S,Houston LL

更新日期：1996-08-01 00:00:00