Abstract:
:Androgen receptor (AR)-associated coregulator 70 (ARA70) is a cytoplasmic protein that has been characterized to have the ability to induce AR transcriptional activity in response to androgens and anti-androgens in prostate cancer cells. AR has been shown to have an important role in the progression of prostate cancer and in normal male reproductive system development. To elucidate the molecular mechanisms and biological relevance of ARA70 to prostrate cancer using a variety of biochemical analyses, the cDNA encoding full length ARA70 was cloned into pET21b vector. Here, we report the refolding and one-step purification of ARA70 from inclusion bodies over-expressed in Escherichia coli. The protein was purified to homogeneity, yielding approximately 60 mg ARA70 from 1L of terrific broth media. Refolding process of ARA70 was monitored using far-UV CD analysis.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Singh VK,Jia Zdoi
10.1016/j.pep.2004.11.002keywords:
subject
Has Abstractpub_date
2005-02-01 00:00:00pages
283-7issue
2eissn
1046-5928issn
1096-0279pii
S1046-5928(04)00379-1journal_volume
39pub_type
杂志文章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
pub_type: 杂志文章
doi:10.1006/prep.1996.0069
更新日期:1996-08-01 00:00:00
abstract::HtrA2 is an apoptosis-activating protein to enhance the apoptotic process by preventing the formation of the IAP-caspase complex, thus freeing caspase to trigger the apoptosis pathway. Here, we presented the full-length sequence of HtrA2 from the black tiger shrimp (PmHtrA2). The full-length PmHtrA2 transcript was 140...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2013.09.012
更新日期:2013-12-01 00:00:00
abstract::Preventing protein aggregation is crucial for various protein studies, and has a large potential for remedy of protein misfolding or aggregates-linked diseases. In this study, we demonstrated the hyper-acidic protein fusion partners, which were previously reported to enhance the soluble expression of aggregation-prone...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2011.06.004
更新日期:2011-11-01 00:00:00
abstract::Influenza A virus displays one of the highest infection rates of all human viruses and therefore represents a severe human health threat associated with an important economical challenge. Influenza matrix protein 2 (M2) is a membrane protein of the viral envelope that forms a proton selective ion channel. Here we repo...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2016.11.001
更新日期:2017-03-01 00:00:00
abstract::Cardiomyopathy-related mutations in human cardiac troponin subunits, including troponin C (hcTnC), troponin I (hcTnI), and troponin T (hcTnT), are well-documented. Recently, it has been recognised that human cardiac troponin (hcTn) is a sophisticated allosteric system. Therefore, the effect of drugs on this protein co...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2012.10.005
更新日期:2013-02-01 00:00:00
abstract::The introduction of an affinity tag offers an attractive approach to isolation of membrane proteins. The type of affinity tag and its positioning in the protein is determined by the desired subsequent experimental uses of the isolated protein. To minimize the risk of interference, membrane proteins may preferentially ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2009.06.005
更新日期:2009-11-01 00:00:00
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
更新日期:2007-12-01 00:00:00
abstract::Interferons (IFNs) are a family of pleiotropic cytokines used for the treatment of various viral infections and cancers. The low-cost production of IFNs with high biological value and the discovery of IFNs with improved properties are important for the treatment of these diseases as well as for understanding the physi...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(03)00187-6
更新日期:2003-10-01 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
pub_type: 杂志文章
doi:10.1006/prep.1995.1034
更新日期:1995-06-01 00:00:00
abstract::Hsp47 is regarded as a collagen-specific chaperone with several suggested roles in collagen biosynthesis under normal and disease conditions. We describe here a procedure for the expression and purification of Hsp47 in Escherichia coli using the IMPACT expression system (New England Biolabs) where the guest gene is fu...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2001.1470
更新日期:2001-10-01 00:00:00
abstract::Over-expression of heterologous proteins in Escherichia coli is commonly hindered by the formation of inclusion bodies. Nevertheless, refolding of proteins in vitro has become an essential requirement in the development of structural genomics (proteomics) and as a means of recovering functional proteins from inclusion...
journal_title:Protein expression and purification
pub_type: 杂志文章,评审
doi:10.1016/j.pep.2007.05.014
更新日期:2007-10-01 00:00:00
abstract::Human paraoxonase (hPON3) is a high density lipoprotein-related glycoprotein with multi-enzymatic properties and antioxidant activity which is proposed to participate in the prevention of low density lipoprotein (LDL) oxidation. In this study, hPON3 gene was amplified from Human Fetal Liver Marathon-Ready cDNA and exp...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.07.021
更新日期:2006-03-01 00:00:00
abstract::PDZ (PSD-95, DiscsLarge, ZO1) domains function in nature as protein binding domains within scaffold and membrane-associated proteins. They comprise ∼90 residues and make specific, high affinity interactions with complementary C-terminal peptide sequences, with other PDZ domains, and with phospholipids. We hypothesized...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2014.02.015
更新日期:2014-06-01 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
pub_type: 杂志文章
doi:10.1006/prep.1999.1120
更新日期:1999-11-01 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
pub_type: 杂志文章
doi:10.1006/prep.1997.0765
更新日期:1997-10-01 00:00:00
abstract::NAD(+)-dependent malic enzyme (NAD-ME) gene from Escherichia coli K12 was inserted into an expression vector pET24b(+) and transformed into E. coli BL21 (DE3). Recombinant NAD-ME was expressed upon IPTG induction, purified with affinity chromatography, and biochemically characterized. The results showed that recombina...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.11.017
更新日期:2007-05-01 00:00:00
abstract::A family of restriction enzyme- and ligation-independent cloning vectors has been developed for producing recombinant His-tagged fusion proteins in Escherichia coli. These are based on pURI2 and pURI3 expression vectors which have been previously used for the successful production of recombinant proteins at the millig...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2010.10.013
更新日期:2011-03-01 00:00:00
abstract::Acyl coenzyme A binding protein (ACBP) has been proposed to transport fatty acyl CoAs intracellularly, facilitating their metabolism. In this study, a new mouse recombinant ACBP was produced by insertion of a histidine (his) tag at the C-terminus to allow efficient purification by Ni-affinity chromatography. The his-t...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2007.11.010
更新日期:2008-04-01 00:00:00
abstract::The leucine aminopeptidase from Aeromonas proteolytica (also known as Vibrio proteolyticus) (AAP) is a metalloenzyme with broad substrate specificity. The open reading frame (ORF) for AAP encodes a 54 kDa enzyme, however, the extracellular enzyme has a molecular weight of 43 kDa. This form of AAP is further processed ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2004.05.004
更新日期:2004-10-01 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
pub_type: 杂志文章
doi:10.1006/prep.1996.0047
更新日期:1996-05-01 00:00:00
abstract::Our earlier studies have demonstrated that the 35 kDa isoform of Translocated promoter region protein (Tpr) of Rattus norvegicus was able to augment c-jun transcription efficiently. Identification of direct targets that may in part downregulate c-jun transcription might prove to be an ideal target to curtail the proli...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2011.01.001
更新日期:2011-05-01 00:00:00
abstract::Production of antibody fragments in heterologous hosts such as Escherichiacoli provides a unique and cost-effective method to develop engineered vectors for tumor targeting. A single-chain Fragment variable (scFv) of the murine monoclonal antibody MAb-B43.13 targeting CA125 in epithelial ovarian cancer was previously ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2014.07.007
更新日期:2014-10-01 00:00:00
abstract::The purpose of this work was to characterize the functions of two sesquiterpene synthase genes from moso bamboo (Phyllostachys edulis). Two novel sesquiterpene synthase genes, belonging to the Tpsa subfamily, were isolated from moso bamboo. MoTPS2 was 1641 bp in length and encoded a protein of 63 kDa, whereas MoTPS6 w...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2015.11.019
更新日期:2016-04-01 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
pub_type: 杂志文章
doi:10.1006/prep.1994.1040
更新日期:1994-06-01 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
pub_type: 杂志文章
doi:10.1006/prep.2000.1340
更新日期:2001-02-01 00:00:00
abstract::The sequence and structure of the target protein exert a marked effect on its soluble expression in Escherichia coli. The effects of the mutation of an amylase isolated from Bacillus licheniformis (BLA) on its soluble expression in E. coli were investigated. A random mutation library of BLA was constructed to screen f...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2015.12.010
更新日期:2016-04-01 00:00:00
abstract::Bovine follicle-stimulating hormone (bFSH) is a pituitary gonadotropin composed of two non-covalently associated polypeptide subunits, which must be glycosylated, folded, and assembled as a heterodimer to be biologically active. Low-level expression of the recombinant bFSH is the factor that limits its usefulness as a...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2007.03.016
更新日期:2007-08-01 00:00:00
abstract::To obtain strong bacterial expression of proteins that seem to be hard to express in bacteria or are highly toxic for bacteria, it is possible to create a palette of similar constructs, differing only by several nucleotides, gradually deleted from the full-length clone by exonuclease III. When a construct is equipped ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2004.12.013
更新日期:2005-05-01 00:00:00
abstract::Botulinum neurotoxin serotype A (BoNT/A) is an extremely potent bacterial protein toxin. The Hc fragment of BoNT/A (AHc) was shown to be non-toxic, antigenic, and capable of eliciting a protective immunity in animals challenged with homologous BoNT. In this study, we synthesized AHc gene by using T4 DNA ligase and PCR...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2009.11.007
更新日期:2010-05-01 00:00:00
abstract::A method is presented to produce large amounts of Bcl-2 and Bcl-x(L), two anti-apoptotic proteins of considerable biomedical interest. Expression constructs were prepared in which the Escherichia coli protein TolAIII, known to promote over expression of soluble product, was added to the N-terminus of Bcl-2 or Bcl-x(L)...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2008.04.005
更新日期:2008-08-01 00:00:00