Abstract：
:Fusion proteins expressed in bacteria are often insoluble or inefficiently purified by standard procedures previously reported to work well for the non-fused proteins. We report here a simple but general procedure that can be used to quickly customize and optimize the purification of milligram quantities of most GST fusion proteins. For each new protein, this procedure determines the optimal conditions for solubilization with detergents in a bacterial lysate, binding to glutathione-agarose beads, and elution with different buffers. This approach was applied to three GST fusion proteins containing large fragments of the Hox transcription factors Lox2, Lox4, and Lox6 that had low solubility and poor elution when purified following published procedures. After optimization, purified proteins were obtained at high yield and successfully used to raise and purify antibodies for the study of the expression patterns of these genes in embryonic tissues.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Mercado-Pimentel ME,Jordan NC,Aisemberg GOdoi
10.1016/s1046-5928(02)00524-7keywords:
subject
Has Abstractpub_date
2002-11-01 00:00:00pages
260-5issue
2eissn
1046-5928issn
1096-0279pii
S1046592802005247journal_volume
26pub_type
杂志文章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
pub_type： 杂志文章
doi：10.1006/prep.1996.0005
更新日期：1996-02-01 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
pub_type： 杂志文章
doi：10.1006/prep.1998.0995
更新日期：1999-02-01 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
pub_type： 杂志文章
doi：10.1016/j.pep.2008.09.008
更新日期：2009-02-01 00:00:00
abstract：:Pseudolysin is the extracellular elastase of Pseudomonas aeruginosa and belongs to the thermolysin-like family of metallopeptidases. Pseudolysin has been identified as a robust drug target and a biotechnologically important enzyme in the tanning industry. Previous attempts to purify active pseudolysin from P. aerugino...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2015.05.005
更新日期：2015-09-01 00:00:00
abstract：:The complement regulatory protein (CRP) of Trypanosoma cruzi is a developmentally regulated glycosylphosphatidylinositol (GPI)-anchored membrane protein that protects the parasite from complement-mediated killing, and is an important virulence determinant of the microorganism. CRP binds human complement components C3b...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(02)00562-4
更新日期：2003-01-01 00:00:00
abstract：:Leishmaniasis is considered by the World Health Organization to be the second most important disease caused by a protozoan parasite. Biochemical and molecular biology studies can help in the understanding of the biology of the Leishmania parasite. All protozoan parasites, including Leishmania, are unable to synthesize...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2006.05.010
更新日期：2006-10-01 00:00:00
abstract：:Insulin-like growth factor binding proteins (IGFBPs) display many functions in humans including regulation of the insulin-like growth factor (IGF) signaling pathway. The various roles of human IGFBPs make them attractive protein candidates in drug discovery. Structural and functional knowledge on human proteins with t...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2014.10.017
更新日期：2015-04-01 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
pub_type： 杂志文章
doi：10.1016/j.pep.2019.105546
更新日期：2020-03-01 00:00:00
abstract：:The gene encoding a novel glucoamylase (GlucaM) from the Corallococcus sp. strain EGB was cloned and heterologous expressed in Escherichia coli BL21(DE3), and the enzymatic characterization of recombinant GlucaM (rGlucaM) was determined in the study. The glucaM had an open reading frame of 1938 bp encoding GlucaM of 6...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2015.06.009
更新日期：2017-01-01 00:00:00
abstract：:Antifreeze proteins (AFPs) enable organisms to survive under freezing or sub-freezing conditions. AFPs have a great potential in the low temperature storage of cells, tissues, organs, and foods. This process will require a large number of recombinant AFPs. In the present study, the recombinant carrot AFP was highly ex...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.01.019
更新日期：2004-06-01 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：:Proteins are essential throughout the biological and biomedical sciences and the purification strategies of proteins of interest have advanced over centuries. Elastin-like polypeptides (ELPs) are compound polymers that have recently been highlighted for their sharp and reversible phase transition property when heated ...
journal_title：Protein expression and purification
pub_type： 杂志文章,评审
doi：10.1016/j.pep.2018.09.006
更新日期：2019-01-01 00:00:00
abstract：:Human granulocyte-macrophage colony stimulating factor (hGM-CSF) is a hematopoietic growth factor. It is widely employed as a therapeutic agent targeting neutropenia in cancer patients undergoing chemotherapy and in patients with AIDS or after bone marrow transplantation. In this study, we constructed the recombinant ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2019.03.010
更新日期：2019-07-01 00:00:00
abstract：:Granulocyte-macrophage colony-stimulating factor (GM-CSF) regulates proliferation, differentiation, and function of hematopoietic progenitor cells. Aside from expansion of hematopoietic cells, GM-CSF has shown efficacy in other diseases, including Crohn's disease. While GM-CSF being clinically used in humans, the abil...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2005.08.014
更新日期：2005-12-01 00:00:00
abstract：:Rotavirus VP8* subunit is the minor trypsin cleavage product of the spike protein VP4, which is the major determinant of the viral infectivity and neutralization. To study the structure-function relationship of this fragment and to obtain type-specific reagents, substantial amounts of this protein are needed. Thus, fu...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2005.09.014
更新日期：2006-04-01 00:00:00
abstract：:Developmentally regulated G-proteins (DRGs) are a highly conserved family of GTP-binding proteins found in archaea, plants, fungi and animals, indicating important roles in fundamental pathways. Their function is poorly understood, but they have been implicated in cell division, proliferation, and growth, as well as s...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2009.05.009
更新日期：2009-10-01 00:00:00
abstract：:ABCA4 is a member of the A subfamily of ATP-binding cassette transporters that consists of large integral membrane proteins implicated in inherited human diseases. ABCA4 assists in the clearance of N-retinylidene-phosphatidylethanolamine, a potentially toxic by-product of the visual cycle formed in photoreceptor cells...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2014.02.010
更新日期：2014-05-01 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
pub_type： 杂志文章
doi：10.1016/1046-5928(92)90005-h
更新日期：1992-08-01 00:00:00
abstract：:Bikunin is a proteoglycan exhibiting broad-spectrum inhibitory activity against serine proteases and could potentially suppress tumor cell invasion and metastasis. Here, we have successfully expressed recombinant human bikunin (rh-bikunin) in the methylotrophic yeast Pichia pastoris and established the purification pr...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2008.03.025
更新日期：2008-08-01 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
pub_type： 杂志文章
doi：10.1006/prep.1998.0910
更新日期：1998-08-01 00:00:00
abstract：:A crude cell extract from yeast Saccharomyces cerevisae was fractionated by affinity chromatography using the leucine tRNA gene as the recognition site. This approach enables the rapid purification of a protein, which retained its full DNA binding capacity during the enrichment procedure. The active fraction contains ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/1046-5928(90)90015-q
更新日期：1990-11-01 00:00:00
abstract：:Amyloid-beta (Aβ) peptide mediates several neurodegenerative diseases. The 42 amino acid (Aβ1-42) is the predominant form of peptide found in the neuritic plaques and has been demonstrated to be neurotoxic in vivo and in vitro. The availability of large quantities of Aβ peptide will help in several biochemical and bio...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2015.05.015
更新日期：2015-10-01 00:00:00
abstract：:Advances in expressed protein ligation (EPL) methods that permit specific introduction of unique modifications into proteins have facilitated protein engineering, structure-function and protein interaction studies. An EPL-generated hybrid exchangeable apolipoprotein has been constructed from recombinant fragments of a...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2007.04.002
更新日期：2007-08-01 00:00:00
abstract：:Achromogenic atypical Aeromonas salmonicida is the causative agent of goldfish ulcer disease. Virulence of this bacterium is associated with the production of a paracrystalline outer membrane A-layer protein. The species-specific structural gene for the monomeric form of A-protein was cloned into a pET-3d plasmid in o...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1999.1054
更新日期：1999-08-01 00:00:00
abstract：:A putative laccase gene (lacG) from Geobacillus sp. JS12 was cloned and expressed as a fusion protein with six histidine residues in Escherichia coli BL21 (DE3) cells, and the protein was primarily found in inclusion bodies. The resulting insoluble proteins were solubilized with 6 M guanidine HCl and refolded using an...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2020.105646
更新日期：2020-09-01 00:00:00
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
更新日期：2018-02-01 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
pub_type： 杂志文章
doi：10.1016/j.pep.2010.06.020
更新日期：2010-11-01 00:00:00
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
更新日期：2013-02-01 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
pub_type： 杂志文章,评审
doi：10.1016/j.pep.2017.08.003
更新日期：2017-12-01 00:00:00
abstract：:Huperzine A (HupA) is a drug used for the treatment of Alzheimer's disease. However, the biosynthesis of this medicinally important compound is not well understood. The HupA biosynthetic pathway is thought to be initiated by the decarboxylation of lysine to form cadaverine, which is then converted to 5-aminopentanal b...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2016.07.013
更新日期：2016-12-01 00:00:00
