当前位置： SCI文献检索 > PROTEIN EXPRESSION AND PURIFICATION期刊下所有文献 > Molecular cloning and functional expression of hemolysin from the sea anemone Actineria villosa.

Molecular cloning and functional expression of hemolysin from the sea anemone Actineria villosa.

Abstract：

:The full-length cDNA that encodes the hemolytic toxin Avt-I, with 226 amino acids, from the venomous sea anemone Actineria villosa has been cloned using the oligo-capping method. The cDNA contains 681bp open reading frame and its predicted amino acid sequences revealed that Avt-I was basic polypeptides without cysteine residues and Arg-Gly-Asp (RGD) motif sequence. The mature Avt-I has a predicted molecular weight of 19.6 kDa and its theoretical isoelectric point is 9.3. The Avt-I revealed 99, 61, 57, and 57% amino acid similarity with hemolytic toxins Pstx20, EqtII, StII, and HmT from Phyllodiscus semoni, Actinia Equina, Stichodactyla helianthus, and Heteractis magnifica, respectively. The characteristic amphiphilic alpha-helix structure was found at the N-terminal region of the mature Avt-I. Recombinant Avt-I (rAvt-I) was expressed in Escherichia coli BL21 (DE3) strain as a biologically active form and purified rAvt-I caused 50% hemolytic activity against 1% sheep erythrocytes at a concentration of 6.3 ng/ml (0.32 nM). M9Y medium led to more than 2-fold increase in rAvt-I yield than cultivation in Luria-Bertani medium.

journal_name

Protein Expr Purif

journal_title

Protein expression and purification

authors

Uechi G,Toma H,Arakawa T,Sato Y

doi

10.1016/j.pep.2004.12.003

keywords:

subject

Has Abstract

pub_date

2005-04-01 00:00:00

pages

379-84

issue

2

eissn

1046-5928

issn

1096-0279

pii

S1046-5928(04)00410-3

journal_volume

40

pub_type

杂志文章

相关文献

PROTEIN EXPRESSION AND PURIFICATION文献大全
  • Cloning, expression, purification, and biochemical characterisation of the FIC motif containing protein of Mycobacterium tuberculosis.

    abstract：:The role of FIC (Filamentation induced by cAMP)(2) domain containing proteins in the regulation of many vital pathways, mostly through the transfer of NMPs from NTPs to specific target proteins (NMPylation), in microorganisms, higher eukaryotes, and plants is emerging. The identity and function of FIC domain containin...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2012.08.020

    authors： Mishra S,Bhagavat R,Chandra N,Vijayarangan N,Rajeswari H,Ajitkumar P

    更新日期：2012-11-01 00:00:00

  • Expression of the Trichoderma reesei tyrosinase 2 in Pichia pastoris: isotopic labeling and physicochemical characterization.

    abstract：:Trichoderma reesei tyrosinase TYR2 has been demonstrated to be able to oxidize various phenolic compounds and also peptide and protein bound tyrosine, and thus is of great interest for different biotechnological applications. In order to understand the reaction mechanism of the enzyme it would be essential to solve it...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2007.04.014

    authors： Westerholm-Parvinen A,Selinheimo E,Boer H,Kalkkinen N,Mattinen M,Saloheimo M

    更新日期：2007-09-01 00:00:00

  • Expression and reconstitution of NF-kappaB from insect cells using a baculovirus vector.

    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

    pub_type： 杂志文章

    doi：10.1006/prep.1996.0670

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

    更新日期：1997-02-01 00:00:00

  • Overexpression and reconstitution of a Rieske iron-sulfur protein from the higher plant.

    abstract：:The iron-sulfur protein subunit, known as the Rieske protein, is one of the central components of the cytochrome b(6)f complex residing in chloroplast and cyanobacterial thylakoid membranes. We have constructed plasmids for overexpression in Escherichia coli of full-length and truncated Rieske (PetC) proteins from the...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/s1046-5928(03)00016-0

    authors： Gubernator B,Seidler A,Rögner M,Szczepaniak A

    更新日期：2003-05-01 00:00:00

  • Expression of the extracellular domain of the human heat-stable enterotoxin receptor in Escherichia coli and generation of neutralizing antibodies.

    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

    pub_type： 杂志文章

    doi：10.1006/prep.1996.0087

    authors： Nandi A,Mathew R,Visweswariah SS

    更新日期：1996-09-01 00:00:00

  • Novel self-cleavage activity of Staphylokinase fusion proteins: An interesting finding and its possible applications.

    abstract：:Staphylokinase (SAK) is reported to have a serine protease domain with no proteolytic activity unlike other plasminogen activators like tissue plasminogen activator (t-PA) and urokinase. A unique protease property of Staphylokinase was observed when SAK was expressed as a fusion protein in inducible Escherichia coli e...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2009.07.011

    authors： Prasad B,Salunkhe SS,Padmanabhan S

    更新日期：2010-02-01 00:00:00

  • Expression and purification of recombinant human tyrosine hydroxylase as a fusion protein in Escherichia coli.

    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

    pub_type： 杂志文章

    doi：10.1016/j.pep.2012.05.007

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

    更新日期：2012-08-01 00:00:00

  • Codon engineering for improved antibody expression in mammalian cells.

    abstract：:While well established in bacterial hosts, the effect of coding sequence variation on protein expression in mammalian systems is poorly characterized outside of viral proteins or proteins from distant phylogenetic families. The potential impact is substantial given the extensive use of mammalian expression systems in ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2007.05.017

    authors： Carton JM,Sauerwald T,Hawley-Nelson P,Morse B,Peffer N,Beck H,Lu J,Cotty A,Amegadzie B,Sweet R

    更新日期：2007-10-01 00:00:00

  • Production of Phosphorylated Ric-8A proteins using protein kinase CK2.

    abstract：:Resistance to Inhibitors of Cholinesterase-8 (Ric-8) proteins are molecular chaperones that fold heterotrimeric G protein α subunits shortly after biosynthesis. Ric-8 proteins also act as test tube guanine nucleotide exchange factors (GEF) that promote Gα subunit GDP for GTP exchange. The GEF and chaperoning activitie...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2018.10.002

    authors： Yu W,Yu M,Papasergi-Scott MM,Tall GG

    更新日期：2019-02-01 00:00:00

  • Human CYP1A1 allelic variants: baculovirus expression and purification, hydrodynamic, spectral, and catalytical properties and their potency in the formation of all-trans-retinoic acid.

    abstract：:Three human cytochrome P450 1A1 (CYP1A1) allelic variants, namely wild-type (CYP1A1.1), CYP1A1.2 (I462V), and CYP1A1.4 (T461N), were expressed as C-terminal His-tagged fusions including a thrombin cleavage site in Spodoptera frugiperda insect cells by baculovirus infection. The variants were expressed with 30-90 nmol ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/s1046-5928(02)00672-1

    authors： Chernogolov A,Behlke J,Schunck WH,Roots I,Schwarz D

    更新日期：2003-04-01 00:00:00

  • Purification and characterization of recombinant Plasmodium falciparum adenylosuccinate synthetase expressed in Escherichia coli.

    abstract：:Most parasitic protozoa lack the de novo purine biosynthetic pathway and rely exclusively on the salvage pathway for their purine nucleotide requirements. Enzymes of the salvage pathway are, therefore, candidate drug targets. We have cloned the Plasmodium falciparum adenylosuccinate synthetase gene. In the parasite, a...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.2001.1610

    authors： Jayalakshmi R,Sumathy K,Balaram H

    更新日期：2002-06-01 00:00:00

  • Immediate-early baculovirus vectors for foreign gene expression in transformed or infected insect cells.

    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

    pub_type： 杂志文章

    doi：10.1006/prep.1996.0092

    authors： Jarvis DL,Weinkauf C,Guarino LA

    更新日期：1996-09-01 00:00:00

  • Purification and partial characterization of acyl carrier protein from Euglena gracilis variety bacillaris.

    abstract：:Acyl carrier protein (ACP) was purified from Euglena gracilis variety bacillaris in yields of about 1 mg/100 g (wet wt) of cells. Antibodies against the purified protein were raised in hens and isolated from eggs. Antibodies raised against Euglena ACP inhibited the Euglena chloroplast nonaggregated fatty acid syntheta...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/1046-5928(91)90072-q

    authors： Williams SG,Worsham LM,Ernst-Fonberg ML

    更新日期：1991-04-01 00:00:00

  • A simple and efficient method for generating high-quality recombinant Mical enzyme for in vitro assays.

    abstract：:We have recently identified a new family of multidomain oxidoreductase (redox) enzymes, the MICALs, that directly regulate the actin cytoskeletal elements necessary for the morphology, motility, and trajectory of cells. Our genetic assays reveal that Mical is both necessary and sufficient for actin organization and ce...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2016.05.008

    authors： Wu H,Hung RJ,Terman JR

    更新日期：2016-11-01 00:00:00

  • Site- and subunit-specific incorporation of unnatural amino acids into HIV-1 reverse transcriptase.

    abstract：:A highly efficient cell-free translation system has been combined with suppressor tRNA technology to substitute nor-Tyr and 3-fluoro-Tyr in place of Tyr183 at the DNA polymerase active site of p66 of human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT). Supplementing the wild-type HIV-1 p51 RT subunit ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2004.07.019

    authors： Klarmann GJ,Eisenhauer BM,Zhang Y,Sitaraman K,Chatterjee DK,Hecht SM,Le Grice SF

    更新日期：2004-11-01 00:00:00

  • The screening of expression and purification conditions for replicative DNA polymerase associated B-subunits, assignment of the exonuclease activity to the C-terminus of archaeal pol D DP1 subunit.

    abstract：:The B-subunits of replicative DNA polymerases belong to the superfamily of calcineurin-like phosphoesterases and are conserved from Archaea to humans. Recently we and others have shown that the B-subunit (DP1) of the archaeal family D DNA polymerase is responsible for proofreading 3'-5' exonuclease activity. The simil...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2005.05.002

    authors： Jokela M,Raki M,Heikkinen K,Sepponen K,Eskelinen A,Syväoja JE

    更新日期：2005-09-01 00:00:00

  • Generation and identification of variants with improved purification yield of Bowman-Birk protease inhibitors carrying protein binding loops.

    abstract：:Replacing the chymotrypsin inhibitory loop of soybean Bowman-Birk inhibitor (sBBI) with a VEGF binding peptide (BBI-AV) significantly reduces the overall purification yield when BBI-AV is produced as a fusion protein in a Bacillussubtilis expression system. The low purification yield is primarily due to a higher fract...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2009.08.006

    authors： Collier KD,Vogtentanz G,Amin NS,Estabrook M,Estell DA,Fox B,Power SD,Rao R,Schmidt BF

    更新日期：2009-12-01 00:00:00

  • A general path for large-scale solubilization of cellular proteins: from membrane receptors to multiprotein complexes.

    abstract：:Expression of recombinant proteins in bacterial or eukaryotic systems often results in aggregation rendering them unavailable for biochemical or structural studies. Protein aggregation is a costly problem for biomedical research. It forces research laboratories and the biomedical industry to search for alternative, mo...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2012.10.007

    authors： Pullara F,Guerrero-Santoro J,Calero M,Zhang Q,Peng Y,Spåhr H,Kornberg GL,Cusimano A,Stevenson HP,Santamaria-Suarez H,Reynolds SL,Brown IS,Monga SP,Van Houten B,Rapić-Otrin V,Calero G,Levine AS

    更新日期：2013-02-01 00:00:00

  • Purification of Fab fragments from a monoclonal antibody papain digest by Gradiflow electrophoresis.

    abstract：:Fab fragments isolated from papain digests of monoclonal antibodies have a wide variety of uses in analytical and in both in vivo and in vitro diagnostic applications. A novel, non-affinity method which uses the Gradiflow to purify Fab fragments from the papain digest of a mouse IgG1 anti-c-myc monoclonal antibody is ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2003.07.005

    authors： Coleman L,Mahler SM

    更新日期：2003-12-01 00:00:00

  • Overexpression of (His)6-tagged human arginase I in Saccharomyces cerevisiae and enzyme purification using metal affinity chromatography.

    abstract：:Arginase (EC 3.5.3.1; L-arginine amidinohydrolase) is a key enzyme of the urea cycle that catalyses the conversion of arginine to ornithine and urea, which is the final cytosolic reaction of urea formation in the mammalian liver. The recombinant strain of the yeast Saccharomyces cerevisiae that is capable of overprodu...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2011.09.001

    authors： Zakalskiy AE,Zakalska OM,Rzhepetskyy YA,Potocka N,Stasyk OV,Horak D,Gonchar MV

    更新日期：2012-01-01 00:00:00

  • Cloning, overexpression, and purification of glucose-6-phosphate dehydrogenase of Pseudomonas aeruginosa.

    abstract：:Glucose-6-phosphate dehydrogenase (G6PDH) (EC 1.1.1.363) plays an important role in the human pathogen Pseudomonas aeruginosa because it generates NADPH, an essential cofactor for several biosynthetic pathways and antioxidant enzymes. P. aeruginosa G6PDH is also a key enzyme in the metabolism of various carbon sources...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2017.10.004

    authors： Acero-Navarro KE,Jiménez-Ramírez M,Villalobos MA,Vargas-Martínez R,Perales-Vela HV,Velasco-García R

    更新日期：2018-02-01 00:00:00

  • Expression, purification and preliminary NMR characterization of isotopically labeled wild-type human heterotrimeric G protein αi1.

    abstract：:Molecular-level investigation of proteins is increasingly important to researchers trying to understand the mechanisms of signal transmission. Heterotrimeric G proteins control the activation of many critical signal transmission cascades and are also implicated in numerous diseases. As part of a longer-term investigat...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2012.06.003

    authors： Maly J,Crowhurst KA

    更新日期：2012-08-01 00:00:00

  • Improved solubility of replication factor C (RFC) Walker A mutants.

    abstract：:Protein insolubility often poses a significant problem during purification protocols and in enzyme assays, especially for eukaryotic proteins expressed in a recombinant bacterial system. The limited solubility of replication factor C (RFC), the clamp loader complex from Saccharomyces cerevisiae, has been previously do...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2012.03.010

    authors： Marzahn MR,Bloom LB

    更新日期：2012-06-01 00:00:00

  • Expression and bioactivity analysis of Staphylococcal enterotoxin M and N.

    abstract：:Staphylococcal enterotoxins (SEs) are powerful superantigens that stimulate non-specific T-cell proliferation produced by Staphylococcus aureus and draw considerable attention as ideal drugs for cancer therapy. The filtrate of S. aureus culture has been used as ampul named Staphylococcal enterotoxin C injection in cli...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2007.08.005

    authors： Pan YQ,Ding D,Li DX,Chen SQ

    更新日期：2007-12-01 00:00:00

  • One-step affinity tag purification of full-length recombinant human AP-1 complexes from bacterial inclusion bodies using a polycistronic expression system.

    abstract：:The AP-1 transcription factor is a dimeric protein complex formed primarily between Jun (c-Jun, JunB, JunD) and Fos (c-Fos, FosB, Fra-1, Fra-2) family members. These distinct AP-1 complexes are expressed in many cell types and modulate target gene expression implicated in cell proliferation, differentiation, and stres...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2008.01.016

    authors： Wang WM,Lee AY,Chiang CM

    更新日期：2008-05-01 00:00:00

  • Expression, subcellular localization, and enzyme activity of a recombinant human extra-cellular superoxide dismutase in tobacco (Nicotiana benthamiana L.).

    abstract：:Human extracellular superoxide dismutase (hEC-SOD) is an enzyme that scavenges reactive oxygen species (ROS). Because of its antioxidant activity, hEC-SOD has been used as a therapeutic protein to treat skin disease and arthritis in mammalian systems. In this study, codon-optimized hEC-SOD was expressed in tobacco (Ni...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2015.11.014

    authors： Park KY,Kim EY,Lee W,Kim TY,Kim WT

    更新日期：2016-03-01 00:00:00

  • An optimized system for expression and purification of secreted bacterial proteins.

    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

    pub_type： 杂志文章

    doi：10.1016/j.pep.2005.09.003

    authors： Geisbrecht BV,Bouyain S,Pop M

    更新日期：2006-03-01 00:00:00

  • Improving storage stability of recombinant organophosphorus hydrolase.

    abstract：:Organophosphorus hydrolase (OPH) is a ∼38kDa enzyme encoded by opd gene of Flavobacterium sp. The enzyme can hydrolyze and inactivate variety of organophosphate (OP)-compounds, including chemical warfare nerve agents. Thus, OPH is a strong candidate for the development of therapeutic intervention against OP-poisoning ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2015.01.012

    authors： Satvik Iyengar AR,Tripathy RK,Bajaj P,Pande AH

    更新日期：2015-07-01 00:00:00

  • Expression of mammalian Rab Escort protein-1 and -2 in yeast Saccharomyces cerevisiae.

    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

    pub_type： 杂志文章

    doi：10.1016/s1046-5928(02)00506-5

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

    更新日期：2002-10-01 00:00:00

  • High-level synthesis of recombinant murine endostatin in Chinese hamster ovary cells.

    abstract：:Endostatin, a carboxy-terminal fragment of collagen XVIII, has been shown to act as an anti-angiogenic agent that specifically inhibits proliferation of endothelial cells and growth of various primary tumors. Here, we describe the expression by Chinese hamster ovary (CHO) cells of murine endostatin and of a tagged-fus...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2004.01.003

    authors： Chura-Chambi RM,Tornieri PH,Spencer PJ,Nascimento PA,Mathor MB,Morganti L

    更新日期：2004-05-01 00:00:00