Expression, purification, and biochemical characterization of enteroaggregative Escherichia coli heat-stable enterotoxin 1.

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：: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：:Chloroplast transformation systems offer unique advantages in biotechnology, including high level of foreign gene expression, maternal inheritance, and polycistronic expression. We studied chloroplast expression of LTK63 (change Ser-->Lys at position 63 in the A subunit) which is the mutant of Escherichia coli heat-la...

journal_title：Protein expression and purification

authors： Kang TJ,Han SC,Kim MY,Kim YS,Yang MS

更新日期：2004-11-01 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：:To investigate the expression and purification of an unstable heterologous protein in Pichia pastoris, the cDNA of H5-lysozyme, a hen egg lysozyme mutant with a hydrophobic pentapeptide (Phe-Phe-Val-Ala-Pro) fused to the carboxyl terminus, was integrated into the genome of P. pastoris. It was found that medium composi...

journal_title：Protein expression and purification

authors： Liu ST,Saito A,Azakami H,Kato A

更新日期：2003-02-01 00:00:00

abstract：:The immunosuppressive drug FK506 binds its targets FK506-binding protein (FKBP) family and modulates cellular processes. Recent studies demonstrated that FK506 shows anti-malaria effects. Newly identified FK506-binding protein 35 from Plasmodium falciparum (PfFKBP35) is assumed to be the molecular target of FK506 in t...

journal_title：Protein expression and purification

authors： Yoon HR,Kang CB,Chia J,Tang K,Yoon HS

更新日期：2007-05-01 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：:Tyrosine hydroxylase is the rate-limiting step in the synthesis of dopamine and is tightly regulated. Previous studies have shown it to be covalently modified and potently inhibited by 3,4-dihydroxyphenylacetaldehyde (DOPAL), an endogenous neurotoxin via dopamine catabolism which is relevant to Parkinson's disease. In...

journal_title：Protein expression and purification

authors： Higgins CA,Vermeer LM,Doorn JA,Roman DL

更新日期：2012-08-01 00:00:00

abstract：:There is no high-resolution structure for the membrane domain of the human erythrocyte anion exchanger, AE1 (Band 3). In this report, we have developed an expression and purification strategy for AE1 to be used in crystallization trials. Saccharomyces cerevisiae strain BJ5457 was transformed with an expression vector ...

journal_title：Protein expression and purification

authors： Bonar P,Casey JR

更新日期：2010-11-01 00:00:00

abstract：:We present a novel efficient procedure for high level purification of human IGFBP-3. Insulin-like growth factor-binding proteins (IGFBPs) are key regulators of insulin-like growth factor mediated signal transduction and thereby can profoundly influence cellular phenotypes. Certain IGFBPs, including IGFBP-3, have also ...

journal_title：Protein expression and purification

authors： Pircher H,Matscheski A,Laich A,Hermann M,Moser B,Viertler HP,Micutkova L,Lindner H,Sarg B,Zwerschke W,Jansen-Dürr P

更新日期：2010-06-01 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：:Splicing factor proline- and glutamine-rich (SFPQ) is an RNA-binding protein, playing significant roles in gene regulation and subnuclear body formation. Our recent serendipitous discovery showed that SFPQ binds zinc directly and forms an infinite polymer that is induced by zinc binding to the protein. The zinc-induce...

journal_title：Protein expression and purification

authors： Lim YW,Lee M

更新日期：2020-07-01 00:00:00

abstract：:The gene from Aeromonas veronii bv. sobria encoding the metallo-beta-lactamase ImiS was subcloned into pET-26b, and ImiS was over-expressed in BL21(DE3) Escherichia coli and purified using SP-Sepharose chromatography. This protocol yielded over 5 mg of ImiS per liter of growth culture under optimum conditions. The bio...

journal_title：Protein expression and purification

authors： Crawford PA,Sharma N,Chandrasekar S,Sigdel T,Walsh TR,Spencer J,Crowder MW

更新日期：2004-08-01 00:00:00

abstract：:Rab GTPases are post-translationally geranylgeranylated on their C-terminal cysteines by Rab geranylgeranyl transferase (RabGGTase) and this modification is essential for their biological activity. Rab Escort Protein (REP) is a molecular chaperone that assists in the prenylation reaction carried out by RabGGTase. Muta...

journal_title：Protein expression and purification

authors： Sidorovitch V,Niculae A,Kan N,Ceacareanu AC,Alexandrov K

更新日期：2002-10-01 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：: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：: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：: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：:Antimicrobial peptides (AMPs) consist of molecules acting on the defense systems of numerous organisms toward tumor and multiple pathogens, such as bacteria, fungi, viruses, and parasites. Compared to traditional antibiotics, AMPs are more stable and have lower propensity for developing resistance through functioning ...

journal_title：Protein expression and purification

authors： Deng T,Ge H,He H,Liu Y,Zhai C,Feng L,Yi L

更新日期：2017-12-01 00:00:00

abstract：:Structural biology places a high demand on proteins both in terms of quality and quantity. Although many protein expression and purification systems have been developed, an efficient and simple system which can be easily adapted is desirable. Here, we report a new system which combines improved expression, solubility ...

journal_title：Protein expression and purification

authors： Liu H,Naismith JH

更新日期：2009-02-01 00:00:00

abstract：:The coding region for arginyl-tRNA synthetase from jack bean (Canavalia ensiformis) has been sequenced and cloned into the bacterial expression vector pET32a. Transformation of BL21 cells and induction with IPTG results in the high level expression of the protein fused N-terminally with thioredoxin and bearing a His-t...

journal_title：Protein expression and purification

authors： Hogg J,Schiefermayr E,Schiltz E,Igloi GL

更新日期：2008-10-01 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：:Cystatin C is a cysteine protease inhibitor produced by a variety of human tissues. The blood concentration of cystatin C depends on the glomerular filtration rate and is an endogenous marker of renal dysfunction. Recombinant cystatin C protein with high immunogenicity is therefore in demand for the diagnostic market....

journal_title：Protein expression and purification

authors： Hayashi M,Iwamoto S,Sato S,Sudo S,Takagi M,Sakai H,Hayakawa T

更新日期：2013-04-01 00:00:00

abstract：:The fluorescent reporter enhanced Green Fluorescent Protein (EGFP) has been used for assaying a wide range of biological activities ranging from gene expression, or localization of target proteins through to intermolecular interactions. However, over-production of this protein in Escherichia coli has resulted in the p...

journal_title：Protein expression and purification

authors： Youell J,Fordham D,Firman K

更新日期：2011-10-01 00:00:00

abstract：:Receptor for activated C-kinase (RACK1) binds to protein kinase C and functions as an anchor for several other cellular components. Most in vitro studies of RACK1 have been carried out with RACK1 fused to a soluble fusion protein partner, such as GST or MBP. Here, we show that fusion complexes may exist as large solub...

journal_title：Protein expression and purification

authors： Bjørndal B,Trave G,Hageberg I,Lillehaug JR,Raae AJ

更新日期：2003-09-01 00:00:00

abstract：:The presence of antibiotic resistance genes in genetically modified bacteria raises a regulatory concern in the production of therapeutic proteins and additionally reduces the number of plasmids available for propagation in a cell. Cre recombinase from bacteriophage P1, involved in Cre/loxP mechanism is one of the wid...

journal_title：Protein expression and purification

authors： D S,Shyam Mohan AH,Rao SN

更新日期：2020-03-01 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：:A hexahistidine tag was fused to the N-terminus of apoaequorin. A suitable vector encoding the fusion protein was constructed and used for transformation of Escherichia coli JM109 cells. Apoaequorin was overexpressed under the control of tac promoter. It was found, however, that most of the protein existed in the form...

journal_title：Protein expression and purification

authors： Glynou K,Ioannou PC,Christopoulos TK

更新日期：2003-02-01 00:00:00

abstract：:Protein asparagine (N)-linked glycosylation is a post-translational modification that occurs in the endoplasmic reticulum; it plays an important role in protein folding, oligomerization, quality control, sorting, and transport. Accordingly, disorders of glycosylation may affect practically every organ system. Dehydrod...

journal_title：Protein expression and purification

authors： Giladi M,Edri I,Goldenberg M,Newman H,Strulovich R,Khananshvili D,Haitin Y,Loewenstein A

更新日期：2017-04-01 00:00:00

abstract：:Escherichia coli tyrosine kinase (Etk) is a membrane bound kinase in gram-negative bacteria that regulates the export of capsular polysaccharides (CPS). The molecular mechanism behind CPS regulation remains unclear, despite access to a crystal structure of the cytoplasmic kinase domain of Etk. In this study, an effici...

journal_title：Protein expression and purification

authors： Chesterman C,Jia Z

更新日期：2016-09-01 00:00:00